Drosophila melanogaster is a highly versatile model organism that has profoundly advanced our understanding of human diseases. With more than 60% of its genes having human homologs, Drosophila provides an invaluable system for modelling a wide range of pathologies, including neurodegenerative disorders, cancer, metabolic diseases, as well as cardiac and muscular conditions. This review highlights key developments in utilizing Drosophila for disease modelling, emphasizing the genetic tools that have transformed research in this field. Technologies such as the GAL4/UAS system, RNA interference (RNAi) and CRISPR-Cas9 have enabled precise genetic manipulation, with CRISPR-Cas9 allowing for the introduction of human disease mutations into orthologous Drosophila genes. These approaches have yielded critical insights into disease mechanisms, identified novel therapeutic targets and facilitated both drug screening and toxicological studies. Articles were selected based on their relevance, impact and contribution to the field, with a particular focus on studies offering innovative perspectives on disease mechanisms or therapeutic strategies. Our findings emphasize the central role of Drosophila in studying complex human diseases, underscoring its genetic similarities to humans and its effectiveness in modelling conditions such as Alzheimer's disease, Parkinson's disease and cancer. This review reaffirms Drosophila's critical role as a model organism, highlighting its potential to drive future research and therapeutic advancements.

The formation of broodstock gametes is closely linked to the yield and quality in aquaculture production, yet molecular mechanisms underlying this process remain insufficiently understood. The noble scallop Chlamys nobilis, an economically significant dioecious bivalve species, serves as an excellent model for studying gametogenesis. In this study, the adult scallops with testis at different developmental stage were chosen for histological examination and transcriptome analysis to dig genes related gonad development. Totally, 2663 DEGs and their set modules significantly related to spermatogenesis were obtained using WGCNA, including 40 candidate genes represented by EVM713. The gene was specifically expressed in the testis. RNA interference (RNAi) of EVM713 led to impaired testis development, marked by sparse sperm cell arrangement, spermatocytes detaching from the follicle wall, and reduced spermatocyte numbers. Meanwhile, 24 h after RNAi, the expression levels of Bax, and Caspase3 significantly increased (P < 0.05), while those of Bcl2, Dmrt2 and Tssk4 were significantly decreased (P < 0.05). These results indicate that EVM713 is essential for spermatogenesis in bivalves, regulating testis development through the modulation of Dmrt2 and Tssk4 expression. This study provides the first evidence of EVM713 function in mollusks, which is conducive to better understanding molecular mechanisms underlying gametogenesis in marine invertebrates.

The field of RNA research has experienced significant changes and is now at the forefront of contemporary drug development. This narrative overview explores the scientific developments and historical turning points in RNA research, emphasising the field's critical significance in the development of novel therapeutics. Important discoveries like antisense oligonucleotides (ASOs), mRNA therapies, and RNA interference (RNAi) have created novel treatment options that can be targeted, such as the ground-breaking mRNA vaccinations against COVID-19. Advances in high-throughput sequencing, single-cell RNA sequencing, and epitranscriptomics have further unravelled the complexity of RNA biology, shedding light on the intricacies of gene regulation and cellular diversity. The integration of computational tools and bioinformatics has propelled the identification of RNA-based biomarkers and the development of RNA therapeutics. Despite significant progress, challenges such as RNA stability, delivery, and off-target effects persist, necessitating continuous innovation and ethical considerations. This review provides a critical insight into the current state and prospects of RNA research, emphasising its transformative potential in drug discovery. By examining the interplay between technological advancements and therapeutic applications, we underscore the promising horizon for RNA-based interventions in treating a myriad of diseases, marking a new era in precision medicine.

RNA therapeutics came to global attention when mRNA-based vaccines provided an answer to the SARS-CoV-2 pandemic. The immense significance of this development notwithstanding, it is important to note that almost a decade prior to the pandemic, RNA drugs had made important inroads toward the amelioration of disease. The first class of RNA therapies to be introduced into clinical use were the antisense oligomers and siRNA drugs which generally induce a therapeutic effect by acting to brake or to modulate mRNA expression. RNA therapeutics is quickly becoming the fourth pillar of pharmacotherapy, and will have broad applications, including for the treatment of cardiovascular disease.

The United States (US) Food and Drug Administration (FDA) has approved several antisense oligomers (ASOs) and siRNA-based drugs to treat disorders associated with cardiovascular disease. In addition, multiple RNA-based drugs are in clinical trials to assess their safety and efficacy in patients with cardiovascular disorders, such as Zodasiran, a siRNA therapy that targets angiopoietin-like protein 3 (ANGPTL3) to reduce LDL cholesterol.

Because of limitless sequence choice; speed of design; and relative ease of synthesis, RNA drugs will be rapidly developed, will have broad applications, and will be generated at lower cost than other drug types. This review aims to highlight RNA therapies for cardiovascular diseases that are approved, and those that are under clinical evaluation.

Locusts have been a major global agricultural pest that poses a serious threat to crop and livestock production. Entomopathogenic fungi (EPF) provide an eco-friendly control method; however, their efficacy usually takes slow and is unstable. To achieve an enhancement of the biocontrol efficacy of Beauveria bassiana (B. bassiana) against locusts, we developed a new strategy by which B. bassiana and nanocarrier-mediated dsRNA are co-applied across the locust cuticle. The nanocarrier star polycation (SPc) effectively delivers Lmidgf4 dsRNA (dsLmidgf4) into the locust, which targets Locusta migratoria imaginal disc growth factor 4 (Lmidgf4). SPc protects dsLmidgf4 from degradation by the hemolymph and enables efficient gene silencing. Furthermore, SPc has no adverse effects on B. bassiana spore germination and growth. Lmidgf4 interference leads to a thinner layer of endocuticle, thus facilitates infection of B. bassiana, and finally reduces the median lethal time of locusts infected with B. bassiana. In conclusion, the combination of B. bassiana and dsRNA/SPc complex overcomes the slow action of fungi, providing a novel strategy for field control of locusts.

In recent years, success has been achieved in treating several eye conditions with oligonucleotide-based therapies. Herein, we outline the experimentation involved in progressing selection and development of a lead therapeutic siRNA for R124H mutation of TGFBI gene which causes Granular Corneal Dystrophy Type 2 (GCD2/Avellino CD). Firstly, a series of siRNA designs, generated by a gene walk across the R124H TGFBI mutation site, were tested and a lead siRNA identified. The lead siRNA was delivered into an immortalised human corneal epithelial cell line to assess on-target efficacy and off-target effects. The in vivo efficacy of the lead R124H TGFBI siRNA, complexed with Bio-Courier technology, silicon stabilized hybrid lipid nanoparticles (sshLNP), was assessed in a mouse model of GCD2 which expressed the human R124H TGFBI transgene. Following topical siRNA application for 5 consecutive days, expression of the R124H mutant TGFBI transgene was measured and shown to be reduced by 22.4 % (± 15.7 %, p < 0.05). We investigated gene expression in the mouse cornea and showed expression of murine Tgfbi was 20-fold lower than TGFBI in human cornea, and expression of the mutant TGFBI transgene was a further 3-fold lower. This estimated 60-fold lower mutant transgene expression may explain the low frequency of corneal deposits observed in this mouse model, limiting its usefulness to test whether siRNA silencing is capable of phenotypic improvement or regression of GCD2/Avellino corneal dystrophy. We assessed WT TGFBI silencing in human primary corneal epithelial cells (PCEC) derived from human corneal limbal biopsy material, which express TGFBI at a similar level to human corneal biopsy. We demonstrated that a single 100 nM siRNA treatment, delivered by the sshLNP to the primary human corneal epithelial cells, gave 26.6 % (± 6.6 %, p < 0.001) reduction in TGFBI mRNA and a 15.4 % (±10.5 %, p < 0.05 %) reduction in TGFBi protein after 48 h. In consideration of the mutant gene expression levels in existing models of GCD2 disease, an ex vivo model of mutation-expressing primary corneal epithelial cells generated from corneal limbal biopsies from GCD2 patients would be more suitable than existing transgenic mouse models for future pre-clinical work in the development of gene silencing therapies for corneal dystrophies.

Effective gene carriers will promote the application of RNA interference (RNAi) technology in future pesticide development. This paper reports a group of novel methionine-based sulfonium lipid compounds (MSLs) and screens their gene delivery abilities in vitro and in vivo. Experiments showed that most MSLs could encapsulate nucleic acids into nanoparticles at an S/P ratio of 4:1, with nanoparticle sizes ranging from 124 to 216 nm and zeta potentials ranging from +27 to 40 mV, and could effectively protect nucleic acids from enzymatic degradation. MSLs successfully mediated the cellular uptake and transfection of nucleic acids in Kc cells and insects. Using dsRNA of CHT10 as the RNAi target, four MSLs were proven to mediate dsRNA interference in Drosophila melanogaster and Ostrinia furnacalis and achieved significant growth inhibition during larval development, eventually leading to pest death. The study demonstrates that MSLs are useful nanocarriers for the development of dsRNA pesticides.

Expression of double-stranded RNAs (dsRNAs) in plants is an emerging strategy to efficiently control insects. RNA interference (RNAi)-mediated pest control takes advantage of double-stranded RNA that can suppress the expression of one or more insect genes that encode key proteins. Virus-induced gene silencing (VIGS) is a useful tool for plant expression of dsRNAs to control pests without altering the plant's genome. Trehalase (TRE) and chitin synthase (CHS) are very important in insects. In this study, we first demonstrated that spraying dsRNAs targeting CHS and TRE increased the mortality rate of the peach aphid Myzus persicae treated with the pathogenic fungus Metarhizium anisopliae. When dsRNAs targeting mpTRE and mpCHS were expressed in plants via VIGS, the expression of mpTRE and mpCHS was reduced in aphids, and their fertility and survival rates were decreased. These results indicate that VIGS-mediated RNA interference is a powerful approach to effectively control aphids, and aphids had a higher mortality rate when M. anisopliae was sprayed.

The green alga Chlamydomonas is an important and versatile model organism for research topics ranging from photosynthesis and metabolism, cilia, and basal bodies to cellular communication and the cellular cycle and is of significant interest for green bioengineering processes. The genome in this unicellular green alga is contained in 17 haploid chromosomes and codes for 16 883 protein coding genes. Functional genomics, as well as biotechnological applications, rely on the ability to remove, add, and change these genes in a controlled and efficient manner. In this review, the history of gene editing in Chlamydomonas is put in the context of the wider developments in genetics to demonstrate how many of the key developments to engineer these algae follow the global trends and the availability of technology. Building on this background, an overview of the state of the art in Chlamydomonas engineering is given, focusing primarily on the practical aspects while giving examples of recent applications. Commonly encountered Chlamydomonas-specific challenges, recent developments, and community resources are presented, and finally, a comprehensive discussion on the emergence and evolution of CRISPR/Cas-based precision gene editing is given. An outline of possible future paths for gene editing based on current global trends in genetic engineering and tools for gene editing is presented.

This study aimed to investigate the effects of recombinant plasmid heat shock protein 47 small interfering ribonucleic acid (HSP47-siRNA) on fibroblast proliferation and collagenous fibre synthesis in the filtering channel after trabeculectomy in rabbit.

The recombinant plasmid HSP47-siRNA was constructed successfully to extract sufficient recombinant plasmids. Furthermore, an animal model of anti-glaucoma surgery in rabbit eyes was created to establish the HSP47-siRNA group, empty vector group, mitomycin C (MMC) group and normal saline group. This study further observed the postoperative eye condition, measured the intraocular pressure and carried out histological and immunohistochemical staining analysis.

At different time points after surgery, there were no significant differences in the comparison of filtering bleb morphology, conjunctival congestion, corneal oedema, inflammation and depth of anterior chamber or lens opacity among the HSP47-siRNA, empty vector, MMC and normal saline groups. Within 7 days after surgery, filtering blebs in all groups were bulged and diffused, with no statistically significant difference (p > 0.05). On day 15 and day 30 after surgery, the upper filtering bleb gradually decreased in the empty vector group and the normal saline group compared with the HSP47-siRNA group (p < 0.01). On day 30 after surgery, flat and pale scar tissues were observed in the surgical area of the empty vector and normal saline groups, whereas functional blebs were still observed in the HSP47-siRNA and MMC groups. Within 7 days after surgery, the intraocular pressure significantly decreased in each group, with a statistically significant difference compared with that before surgery (p < 0.01).

HSP47-siRNA exhibits application potential concerning its effects on anti-scarring after glaucoma filtration surgery in rabbit.

Resistance to anthelmintics, particularly the macrocyclic lactone ivermectin (IVM), presents a substantial global challenge for parasite control. We found that the functional loss of an evolutionarily conserved E3 ubiquitin ligase, UBR-1, leads to IVM resistance in Caenorhabditis elegans. Multiple IVM-inhibiting activities, including viability, body size, pharyngeal pumping, and locomotion, were significantly ameliorated in various ubr-1 mutants. Interestingly, exogenous application of glutamate induces IVM resistance in wild-type animals. The sensitivity of all IVM-affected phenotypes of ubr-1 is restored by eliminating proteins associated with glutamate metabolism or signaling: GOT-1, a transaminase that converts aspartate to glutamate, and EAT-4, a vesicular glutamate transporter. We demonstrated that IVM-targeted GluCls (glutamate-gated chloride channels) are downregulated and that the IVM-mediated inhibition of serotonin-activated pharynx Ca2+ activity is diminished in ubr-1. Additionally, enhancing glutamate uptake in ubr-1 mutants through ceftriaxone completely restored their IVM sensitivity. Therefore, UBR-1 deficiency-mediated aberrant glutamate signaling leads to ivermectin resistance in C. elegans.

Small interfering RNAs (siRNAs), generated by Dicer proteins, play a pivotal role in antiviral immunity in eukaryotes. Dicer proteins also produce microRNAs (miRNAs), a class of endogenous small non-coding RNAs that regulate essential cellular functions through post-transcriptional mechanisms. In plants and insects, multiple Dicer proteins are produced and deployed to separately manage the biogenesis of antiviral siRNAs and miRNAs. This separation ensures that viral infections, especially the production of viral RNAi suppressors, do not severely compromise host growth or development. In contrast, nematode worms, such as Caenorhabditis elegans, rely on a single Dicer protein to produce both types of small RNAs. Probably as a strategy to mitigate the potential disruption of miRNA production by viral infections, nematodes have evolved distinct strategies for generating primary and secondary siRNAs for antiviral defense. This review explores the shared and unique features of siRNA-mediated antiviral immunity in Caenorhabditis elegans, shedding light on the specialized adaptations that enable robust antiviral defenses without compromising miRNA-mediated function.

Over the past two decades, many double-stranded (ds)-RNAs have been synthesized to silence target genes for exploration of gene functions in pests. Some of these dsRNAs are lethal to pests, leading to a new category of pesticides. The generation of these environmentally friendly pesticides requires precise in silico design of dsRNA molecules that target pests but not non-pest organisms. Current efforts in dsRNA design focus mainly on the analysis of the target gene sequence, lacking comprehensive analysis of all transcripts of the whole transcriptome per given species, causing low efficiency and imprecise dsRNA target exploration. To address these limitations, we created the dsRNAEngineer online platform (https://dsrna-engineer.cn), which allows comprehensive and rational dsRNA design, incorporating hundreds of pest and non-pest transcriptomes. Developed functionalities include screen-target (screen conserved genes for cotargets of various pest species), on-target, off-target, and multi-target to generate optimal dsRNA for precise pest control.

The instability of double-stranded RNA (dsRNA) restricts the application of RNA interference (RNAi) technology in agricultural pest management. Various types of nanocarriers have been developed and employed for the stable delivery of dsRNA. Nonetheless, it remains unclear which type of nanomaterial could deliver dsRNA stably and efficiently for gene knockdown in Locusta migratoria. In this study, we evaluated the ability of three biocompatible and low-toxicity inorganic nanomaterials-polyethylenimine (PEI)-functionalized single-walled carbon nanotube (PEI-SWNT), polyethylenimine-functionalized carbon quantum dots (PEI-CQDs), and layered double hydroxide (LDH)-to bind and stabilize dsRNA. The results revealed that, compared to PEI-CQDs and LDH, PEI-SWNT more effectively protected dsRNA from degradation in locust gut fluids, across various temperatures, and under different pH conditions. Furthermore, we investigated the efficacy of PEI-SWNT/dsRNA complexes in suppressing endogenous genes in locusts through both injection and oral administration methods. Compared to bare dsRNA, PEI-SWNT/dsRNA complexes enhanced RNAi efficiency by up to 46.0 % and increased mortality by up to 39.0 %. Moderate levels of PEI-SWNT could improve the germination rate of wheat, while not affecting leaf growth in the short term. To our knowledge, this study is the first to apply PEI-SWNT inorganic nanomaterials in insects, which provides a foundational basis and compelling evidence for the development of nanomaterial-based nucleic acid pesticides.

Nucleic acid-based therapeutics have become a key pillar of the 'third wave' of modern medicine, following the eras of small molecule inhibitors and antibody drugs. Their rapid progress is heavily dependent on delivery technologies, with the development of N-acetylgalactosamine (GalNAc) conjugates marking a breakthrough in targeting liver diseases. This technology has gained significant attention for its role in addressing chronic conditions like chronic hepatitis B (CHB) and nonalcoholic steatohepatitis (NASH), which are challenging to treat with conventional methods.

This review explores the origins, mechanisms, and advantages of GalNAc-siRNA delivery systems, highlighting their ability to target hepatocytes via the asialoglycoprotein receptor (ASGPR). The literature reviewed covers preclinical and clinical advancements, particularly in CHB and NASH. Key developments in stabilization chemistry and conjugation technologies are examined, emphasizing their impact on enhancing therapeutic efficacy and patient compliance.

GalNAc-siRNA technology represents a transformative advancement in RNA interference (RNAi) therapies, addressing unmet needs in liver-targeted diseases. While significant progress has been made, challenges remain, including restricted targeting scope and scalability concerns. Continued innovation is expected to expand applications, improve delivery efficiency, and overcome limitations, establishing GalNAc-siRNA as a cornerstone for future nucleic acid-based treatments.

Syntaxin 1A (Syx1A) has diverse and indispensable functions in animals. Previous studies have mainly focused on the roles of Syx1A in Drosophila, and so how Syx1A operates during the development of other insects remains poorly understood. This study investigated whether disrupting LmSyx1A using RNA interference (RNAi) affects the growth and development of Locusta migratoria. LmSyx1A was expressed in all tissues tested, with the highest expression observed in the fat body. After 5th-instar nymphs were injected with double-stranded LmSyx1A (dsLmSyx1A), none of the nymphs were able to molt normally and all eventually died. The silencing of LmSyx1A resulted in the cessation of feeding, body weight loss, and atrophy of the midgut and gastric cecum in locusts. Hematoxylin and eosin (H&E) staining showed that the columnar cells in the midgut were severely damaged, with microvilli defects visible in dsLmSyx1A-injected nymphs. Secretory vesicles were observed with transmission electron microscopy (TEM). In addition, reverse transcription quantitative polymerase chain reaction (RT-qPCR) further indicates that silencing LmSyx1A repressed the expression of genes involved in the insulin/mammalian target of rapamycin (mTOR)-associated nutritional pathway. Taken together, these results suggest that LmSyx1A significantly affects the midgut cell layer of locust nymphs, which was partially associated with the downregulation of the insulin/mTOR-associated nutritional pathway. Thus, we argue that LmSyx1A is a suitable target for developing dsRNA-based biological pesticides for managing L. migratoria.

Glyceraldehyde-3-phosphate dehydrogenase 2 (GAPDH2) plays a vital role in cell growth, stress responses, and various cellular processes in organisms. However, its functional characterization in cyanobacteria, particularly in Synechocystis sp. PCC 6803, remains largely unexplored, especially concerning its overexpression and RNA interference (RNAi) via double-stranded RNA (dsRNA). This study aimed to investigate the biological role of GAPDH2 in Synechocystis sp. PCC 6803 by cloning its complete coding sequence (SyGAPDH2). The SyGAPDH2 protein comprises 350 amino acids with a molecular weight of 86.480 kDa and an isoelectric point of 5.03. The sequence alignment analysis revealed two conserved domains: NADH (Nicotinamide Adenine Dinucleotide)-quinone oxidoreductase subunit NuoI and NADH-ubiquinone/plastoquinone oxidoreductase chain 6. Similarly, Phylogenetic analysis demonstrated high sequence similarity of 96 % and 94 % with Coliform (Gammaproteobacteria bacterium), respectively. We further explored the functional significance of SyGAPDH2 through overexpression using the PpsbAII+SyGAPDH2 vector and double stranded RNA (dsRNA)-mediated silencing with dsGAPDH2. Overexpression significantly enhanced cell growth, while dsRNA-mediated suppression resulted in reduced cell proliferation, with effects observed 12 h post-treatment and persisting up to 36 h. These findings emphasize the essential regulatory role of SyGAPDH2 in cellular development and stress response. This study contributes to our understanding of GAPDH2 functional importance in cyanobacteria, providing a foundation for future investigations into its subcellular localization, additional functional roles, and broader regulatory mechanisms within cyanobacterial cellular processes.

Begomoviruses are a group of single stranded DNA plant viruses exclusively transmitted by the sweet potato whitefly Bemisia tabaci in a persistent, circulative manner. After acquisition from plant phloem, this group of viruses circulate and are retained within the whitefly, interacting with tissues, cells and molecular pathways for maintaining the safety of the infective intact virions, by exploiting cellular mechanisms and avoiding degradation by the insect immune responses. During retention, the virions are internalized in the midgut cells, exit and spend hours-days in the hemolymph and cross into salivary gland cells, before transmission. Destroying this group of viruses by the insect immune system seems inefficient for the most part, by examining their very efficient transmission. Thus, within the various sites along the transmission pathway especially in the midgut, it is thought that the immune system with its various layers is activated for avoiding the damage caused by the viruses on one hand, and for ensuring their safe circulation and transmission on the other hand. Begomoviruses have evolved mechanisms for counteracting and exploiting the activated immune system for their safe translocation within the whitefly. In this review, we discuss the various levels of immunity activated against begomoviruses in B. tabaci, taking other pathogen-vector systems as examples and reflecting relevant components on the interactions between B. tabaci and Begomoviruses.

The centrosome is the microtubule-organizing center and a crucial part of cell division. Centrosomal RNAs (cnRNAs) have been reported to enable precise spatiotemporal control of gene expression during cell division in many species. Whether and how cnRNAs exist in C. elegans are unclear. Here, using the nuclear RNAi Argonaute protein NRDE-3 as a reporter, we observed potential peri-centrosome localized small interfering (si)RNAs in C. elegans. NRDE-3 was previously shown to associate with pre-mRNAs and pre-rRNAs via a process involving the presence of complementary siRNAs. We generated a GFP-NRDE-3 knock-in transgene through CRISPR/Cas9 technology and observed that NRDE-3 formed peri-centrosomal foci neighboring the tubulin protein TBB-2, other centriole proteins and pericentriolar material (PCM) components in C. elegans embryos. The peri-centrosomal accumulation of NRDE-3 depends on RNA-dependent RNA polymerase (RdRP)-synthesized 22G siRNAs and the PAZ domain of NRDE-3, which is essential for siRNA binding. Mutation of eri-1, ergo-1, or drh-3 significantly increased the percentage of pericentrosome-enriched NRDE-3. At the metaphase of the cell cycle, NRDE-3 was enriched in both the peri-centrosomal region and the spindle. Moreover, the integrity of centriole proteins and pericentriolar material (PCM) components is also required for the peri-centrosomal accumulation of NRDE-3. Therefore, we concluded that siRNAs could accumulate in the pericentrosomal region in C. elegans and suggested that the peri-centrosomal region may also be a platform for RNAi-mediated gene regulation.

Cancer is a rising issue worldwide, and numerous studies have focused on understanding the underlying reasons for its occurrence and finding proper ways to defeat it. By applying technological advances, researchers are continuously uncovering and updating treatments in cancer therapy. Their vast functions in the regulation of cell growth and proliferation and their significant role in the progression of diseases, including cancer. This review provides a comprehensive analysis of ncRNAs in breast cancer, focusing on long non-coding RNAs such as HOTAIR, MALAT1, and NEAT1, as well as microRNAs such as miR-21, miR-221/222, and miR-155. These ncRNAs are pivotal in regulating cell proliferation, metastasis, drug resistance, and apoptosis. Additionally, we discuss experimental approaches that are useful for studying them and highlight the advantages and challenges of each method. We then explain the results of these clinical trials and offer insights for future studies by discussing major existing gaps. On the basis of an extensive number of studies, this review provides valuable insights into the potential of ncRNAs in cancer therapy. Key findings show that even though the functions of ncRNAs are vast and undeniable in cancer, there are still complications associated with their therapeutic use. Moreover, there is an absence of sufficient experiments regarding their application in mouse models, which is an area to work on. By emphasizing the crucial role of ncRNAs, this review underscores the need for innovative approaches and further studies to explore their potential in cancer therapy.

Transthyretin amyloid cardiomyopathy (ATTR-CM) is a progressive form of heart failure caused by myocardial tissue infiltration with fibrillar amyloid deposits. ATTR-CM has been traditionally underrecognized and regarded by clinicians as a challenging condition to manage, owing to limited availability of effective screening methods, diagnostic testing, and therapeutic options. More recently, multiple clinical trials have emerged evaluating the efficacy of novel pharmacologic therapies which target amyloid generation and pre-existing amyloid deposits. Results reveal robust treatment benefits in function and survival, offering clinicians and patients new therapeutics which alter the clinical trajectory of ATTR-CM. Importantly, the benefits of treatment with these therapies appear to be more pronounced when initiated at an early stage of disease. As a result, a renewed interest in the early detection of ATTR-CM has developed, with efforts currently underway to promote increased disease awareness and enhance diagnosis through standardized screening algorithms and advanced imaging techniques. This review will provide an in-depth description of the advancements in ATTR-CM screening, diagnosis, and treatment that are currently available for implementation in routine care. Furthermore, we highlight several investigational modalities on the horizon for ATTR-CM with a particular focus on their potential roles in future clinical practice.

Nucleic acid-based therapeutics have the ability to tackle a wide range of diseases and stress tolerance that present significant obstacles for conventional approaches in agriculture. RNA-based medicines have become a promising approach, using nanoformulation treatments to specifically target certain diseases. Nanoformulations offer numerous benefits in comparison to alternative treatment methods, such as precise administration, minimal toxicity, and medication loading compatibility due to their bioactivity. There are a variety of nanoformulations available today, such as liposomes, polymeric nanoparticles (NPs), magnetic NPs, nanogels, and solid lipid nanoparticles (SLNs). RNA-based therapy employs intracellular gene nanoparticles containing messenger RNA (mRNA), which play an important role in stress management and pest as well as disease control. The adoption of mRNA-based technology paves the way for future technological progress. This review focuses on elucidating the process underlying the development of RNA interference (RNAi) and the diverse array of nanocarriers employed for the transportation of RNAi. Currently, this technique is being employed in the field of crop protection to combat diseases, pests, and environmental stress. The article highlights the benefits of RNAi mediated nanoformulations and discusses the significant obstacles that must be overcome to improve the viability of this technology for future applications.

Although small interfering RNA (siRNA) holds immense promise for treating genetic diseases and cancers, its clinical application is constrained by instability, cellular uptake barriers, and inefficient cytosolic delivery, underscoring the need for effective delivery systems. Therefore, this study focuses on screening novel T-shaped lipo-xenopeptide (XP) nanocarriers for siRNA polyplex formulation, integrating two single succinoyl-tetraethylene pentamine (Stp) units for electrostatic interaction and tyrosine tripeptides (Y3) for aromatic stabilization, along with structural modifications such as the addition of histidine (H) with or without terminal cysteines (C), and the incorporation of various fatty acids (FAs). A systematic evaluation of siRNA binding, nanoparticle stability, and gene silencing efficiency in multiple cell lines illustrated that the novel Stp1-HC lipo-XPs carriers outperform their Stp2-HC analogs, despite having fewer cationizable Stp units. This advantage stems from increased fatty acid, Y3, and C density, which compensates for reduced electrostatic interactions. The presence of H in combination with unsaturated FAs significantly improved the functional siRNA delivery. Our findings highlight the complex interplay of electrostatic, hydrophobic, covalent, hydrogen-bonded, and aromatic interactions to achieve efficient siRNA delivery, which is best-balanced in the oleic acid-containing Stp1-HC/OleA lipo-XP, exceeding the previously best standard carrier Stp2-HC/OleA in efficiency.

RNA interference (RNAi) has been shown to be relatively effective in coleopteran insects, with limited exploration into the molecular mechanisms that underlie this effectiveness. This study specifically examines the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata (Hvig), known for its high RNAi efficiency. Here, we utilized RNAi and CRISPR/Cas9 techniques to identify and validate the genes involved in the RNAi pathway that enhance RNAi efficacy in Hvig. We identified a total of 15 potential genes within the RNAi pathway that may impact RNAi efficiency. The bioassay results showed that only knockdown of HvStaufenC in the 3rd instar larvae could block the abnormal body color phenotype and lethality induced by the subsequent silencing of the two marker genes, HvTH (tyrosine hydroxylase) and HvABCH1 (ATP-binding cassette H transporter gene), respectively. Additionally, successful CRISPR/Cas9-mediated knockout of HvStaufenC led to the generation of stable, heritable mutants that exhibited insensitivity to RNAi, displaying no response to RNAi targeting HvTH and HvABCH1. Compared to the wild-type strain, the HvStaufenC knockout (HvStaufenCKO) mutant females demonstrated a 42 % decrease in oviposition rate and a 41.3 % reduction in egg hatchability. This study demonstrates that HvStaufenC gene is crucial for the RNAi efficiency of Hvig and offers new evidence into the RNAi mechanisms in coleopteran species.

Lipid nanoparticles (LNP) are a safe and effective messenger RNA (mRNA) delivery system for vaccine applications, as shown by the COVID-19 mRNA vaccines. One of the main challenges faced during the development of these vaccines is the production of new and versatile LNP formulations capable of efficient encapsulation and delivery to cells in vivo. This study aimed to develop a new mRNA vaccine formulation that could potentially be used against existing diseases as well as those caused by pathogens that emerge every year.

Using firefly luciferase (Luc) as a reporter mRNA, we evaluated the physical-chemical properties, stability, and biodistribution of an LNP-mRNA formulation produced using a novel lipid composition and a microfluidic organic-aqueous precipitation method. Using mRNAs encoding a dengue virus or a Leishmania infantum antigen, we evaluated the immunogenicity of LNP-mRNA formulations and compared them with the immunization with the corresponding recombinant protein or plasmid-encoded antigens. For all tested LNP-mRNAs, mRNA encapsulation efficiency was higher than 85%, their diameter was around 100 nm, and their polydispersity index was less than 0.3. Following an intramuscular injection of 10 µg of the LNP-Luc formulation in mice, we detected luciferase activity in the injection site, as well as in the liver and spleen, as early as 6 h post-administration. LNPs containing mRNA encoding virus and parasite antigens were highly immunogenic, as shown by levels of antigen-specific IgG antibody as well as IFN-γ production by splenocytes of immunized animals that were similar to the levels that resulted from immunization with the corresponding recombinant protein or plasmid DNA.

Altogether, these results indicate that these novel LNP-mRNA formulations are highly immunogenic and may be used as novel vaccine candidates for different infectious diseases.

We review empirical methods that can be used to provide physical descriptions of dynamic cellular processes during development and disease. Our focus will be nonspatial descriptions and the inference of underlying interaction networks including cell-state lineages, gene regulatory networks, and molecular interactions in living cells. Our overarching questions are: How much can we learn from just observing? To what degree is it possible to infer causal and/or precise mathematical relationships from observations? We restrict ourselves to data sets arising from only observations, or experiments in which minimal perturbations have taken place to facilitate observation of the systems as they naturally occur. We discuss analysis perspectives in order from those offering the least descriptive power but requiring the least assumptions such as statistical associations. We end with those that are most descriptive, but require stricter assumptions and more previous knowledge of the systems such as causal inference and dynamical systems approaches. We hope to provide and encourage the use of a wide array of options for quantitative cell biologists to learn as much as possible from their observations at all stages of understanding of their system of interest. Finally, we provide our own recipe of how to empirically determine quantitative relationships and growth laws from live-cell microscopy data, the resultant predictions of which can then be verified with perturbation experiments. We also include an extended supplement that describes further inference algorithms and theory for the interested reader.

Storage of genetic information in DNA occurs through a unique ordering of canonical base pairs. However, this would not be possible in the absence of the sugar-phosphate backbone which is essential for duplex formation. While over a hundred nucleobase modifications have been identified (mainly in RNA), Nature is rather conservative when it comes to alterations at the level of the (deoxy)ribose sugar moiety. This trend is not reflected in synthetic analogues of nucleic acids where modifications of the sugar entity is commonplace to improve the properties of DNA and RNA. In this review article, we describe the main incentives behind sugar modifications in nucleic acids and we highlight recent progress in this field with a particular emphasis on therapeutic applications, the development of xeno-nucleic acids (XNAs), and on interrogating nucleic acid etiology. We also describe recent strategies to conjugate carbohydrates and oligosaccharides to oligonucleotides since this represents a particularly powerful strategy to improve the therapeutic index of oligonucleotide drugs. The advent of glycoRNAs combined with progress in nucleic acid and carbohydrate chemistry, protein engineering, and delivery methods will undoubtedly yield more potent sugar-modified nucleic acids for therapeutic, biotechnological, and synthetic biology applications.

Epigenetics influence and are influenced by the impact of social and environmental challenges on biological outcomes. Therefore, pinpointing epigenetic factors associated with social adversity and traumatic stress enables understanding of the mechanisms underlying vulnerability and resilience. We hypothesized that micro-RNAs (miRNAs) expression may be associated with post-traumatic stress disorder symptom severity (i.e., PTSS) following exposure to social adversity. To test this hypothesis, we leveraged blood-derived RNA samples (n=632) and social adversity data from 483 unique participants in the Detroit Neighborhood Health Study, a community-based, prospective cohort of predominantly African Americans. Results identified 86 miRNAs that are associated with social adversities (financial difficulties, perceived discrimination, cumulative trauma) and PTSS. These miRNAs are primarily involved in the immune response, brain and neural function, as well as cell cycle and differentiation, and 22 (25%) have previously been associated with conditions related to PTSD, including traumatic brain injury and stress response. Our findings offer a fresh perspective on understanding the epigenetic role of miRNA in the interaction between social adversity and traumatic stress.

Non-alcoholic fatty liver disease (NAFLD) is a prevalent metabolic liver disorder worldwide, and effective therapeutic strategies for its treatment remains limited. In this article, we introduced Glipo-siRubi, a hepatocytes-targeting RNA interference (RNAi) nanoliposome for suppression of Rubicon expression, aiming to achieve precise regulation of autophagy in NAFLD. Autophagy activation induced by Rubicon suppression resulted in reduced endoplasmic reticulum stress and intracellular lipid accumulation in vitro. Moreover, Glipo-siRubi administration exhibited remarkable therapeutic efficacy, characterized by decreased liver lipid accumulation, ameliorated histopathology and improved insulin sensitivity in mice with western diet, indicating its notable potential against NAFLD. By inducing autophagy activation, the hepatocytes-targeting Glipo-siRubi provided a promising method for NAFLD treatment, addressing the limitations of current approaches. Our study highlighted the significance of Rubicon-specific suppression in NAFLD treatment, offering a specific, safe, and efficient approach to mitigate NAFLD.

Nucleic acid drugs utilize DNA or RNA molecules to modulate abnormal gene expression or protein translation in cells, enabling precise treatment for specific conditions. In recent years, nucleic acid drugs have demonstrated tremendous potential in vaccine development and treating genetic disorders. Currently, the primary carriers for clinically approved nucleic acid therapies include lipid nanoparticles and viral vectors. Beyond that, metal-organic frameworks (MOFs) are highly ordered, porous nanomaterials formed through the self-assembly of metal ions and organic ligands via coordination bonds. Their porosity structure offers great loading efficiency, stability, tunability, and biocompatibility, making them an attractive option for nucleic acid delivery. Given the research on MOFs as nucleic acid carriers has garnered significant attention in recent years, this review provides an overview of the therapeutic strategies and advancements in MOF-mediated nucleic acid delivery. The unique properties of various MOF carriers are introduced, and different approaches for nucleic acid loading are parallelly compared. Moreover, a systematic classification based on the type of nucleic acid cargo loaded in MOFs and corresponding applications is thoroughly described. This summary outlines the unique mechanisms through MOFs enhance nucleic acid delivery and emphasizes their substantial impact on therapeutic efficacy. In addition, the utilization of MOF-mediated nucleic acid treatment in combination with other therapies against malignant tumors is discussed in particular. Finally, an outlook on the challenges and potential opportunities of this technology in future translational production and clinical implementation is presented and explored.

Juvenile hormone (JH) is important to maintain insect larval status; however, its cell membrane receptor has not been identified. Using the lepidopteran insect Helicoverpa armigera (cotton bollworm), a serious agricultural pest, as a model, we determined that receptor tyrosine kinases (RTKs) cadherin 96ca (CAD96CA) and fibroblast growth factor receptor homologue (FGFR1) function as JH cell membrane receptors by their roles in JH-regulated gene expression, larval status maintaining, rapid intracellular calcium increase, phosphorylation of JH intracellular receptor MET1 and cofactor Taiman, and high affinity to JH III. Gene knockout of Cad96ca and Fgfr1 by CRISPR/Cas9 in embryo and knockdown in various insect cells, and overexpression of CAD96CA and FGFR1 in mammalian HEK-293T cells all supported CAD96CA and FGFR1 transmitting JH signal as JH cell membrane receptors.

Corneal fibrosis, a severe complication linked to ocular injuries and post-surgery, lacks effective treatment. Hydrogels are regarded as one of the most promising biomaterials, particularly in the context of corneal wound treatment, where they have attracted considerable attention. Synthetic protein hydrogels are of particular interest due to their biocompatibility, biodegradability, capacity to mitigate induced tissue inflammatory responses, and their editable and modular integrative properties. Accordingly, the present study was designed to create a mechanically stable 4XT recombinant protein based on the mechanism of corneal fibrosis. A bio-synthetic protein gel scaffold incorporating cerium oxide nanoparticles (CeONs) with reactive oxygen species (ROS) scavenging capabilities and siRNA that inhibits transforming growth factor beta 1 (TGF-β1) protein expression was constructed using 4XT as a matrix. This resulted in a composite synthetic protein hydrogel treatment system. This system is capable of achieving in situ curing in the corneal defect area, effectively promoting the repair of corneal wounds in mice while also inhibiting the progression of corneal fibrosis. By combining the programmability and controllability of synthetic protein hydrogels with therapeutic approaches targeting wound mechanisms, it is possible to achieve scarless healing of corneal wounds, thereby providing valuable insights for wound management.

RNA interference (RNAi) is a conserved pathway that utilizes Argonaute proteins and their associated small RNAs to exert gene regulatory function on complementary transcripts. While the majority of germline-expressed RNAi proteins reside in perinuclear germ granules, it is unknown whether and how RNAi pathways are spatially organized in other cell types. Here, we find that the small RNA biogenesis machinery is spatially and temporally organized during Caenorhabditis elegans embryogenesis. Specifically, the RNAi factor, SIMR-1, forms visible concentrates during mid-embryogenesis that contain an RNA-dependent RNA polymerase, a poly-UG polymerase, and the unloaded nuclear Argonaute protein, NRDE-3. Curiously, coincident with the appearance of the SIMR granules, the small RNAs bound to NRDE-3 switch from predominantly CSR-class 22G-RNAs to ERGO-dependent 22G-RNAs. NRDE-3 binds ERGO-dependent 22G-RNAs in the somatic cells of larvae and adults to silence ERGO-target genes; here we further demonstrate that NRDE-3-bound, CSR-class 22G-RNAs repress transcription in oocytes. Thus, our study defines two separable roles for NRDE-3, targeting germline-expressed genes during oogenesis to promote global transcriptional repression, and switching during embryogenesis to repress recently duplicated genes and retrotransposons in somatic cells, highlighting the plasticity of Argonaute proteins and the need for more precise temporal characterization of Argonaute-small RNA interactions.

RNA interference through small interfering RNA (siRNA) has shown great promise as a potential cancer treatment strategy in recent years. However, the delivery of siRNA to target cancer cells efficiently remains a significant challenge. This review aims to highlight the recent advances in nanotechnology-enabled siRNA delivery for cancer treatment, bridging the gap between bench research and clinical application. A comprehensive literature search was conducted to identify recent studies focused on the utilization of nanotechnology for siRNA delivery in cancer treatment. Key databases, including PubMed, Scopus, and Web of Science, were used, and relevant articles were screened. Several nanotechnology-based platforms for siRNA delivery have emerged in recent years, providing enhanced selectivity, improved stability, and controlled release profiles. The primary types of nanocarriers discussed include lipid-based nanoparticles, inorganic nanoparticles, polymeric nanoparticles, and exosomes. Nanotechnology-based siRNA delivery systems represent a promising avenue for cancer treatment. Although significant progress has been made in preclinical studies, translating these findings to clinical applications poses several challenges, including scale-up production, safety, and targeted delivery. Nevertheless, the recent developments in this field hold great promise in revolutionizing cancer therapy, providing hope for more effective and personalized treatment options in the future.

Biomolecular condensates, such as stress and germ granules, often contain subcompartments. For instance, the Caenorhabditis elegans germ granule, which localizes near the outer nuclear membrane of germ cell nuclei, is composed of at least four ordered compartments, each housing distinct sets of proteins and RNAs. How these compartments form and why they are spatially ordered remains poorly understood. Here, we show that the conserved DEAD-box RNA helicase DDX-19 defines another compartment of the larger C. elegans germ granule, which we term the D compartment. The D compartment exhibits properties of a liquid condensate and forms between the outer nuclear pore filament and other compartments of the germ granule. Two nuclear pore proteins, NPP-14 and GLEL-1, are required for its formation, suggesting that the D compartment localizes adjacent to the outer nuclear membrane through interactions with the nuclear pore. The loss of DDX-19, NPP-14 or GLEL-1 leads to functional defects, including aberrant formation of the other four germ granule compartments, a loss of germline immortality and dysregulation of small RNA-based transgenerational epigenetic inheritance programs. Hence, we propose that a function of the D compartment is to anchor larger germ granules to nuclear pores, enabling germ granule compartmentalization and promoting transgenerational RNA surveillance.

Transcriptional profiling of stem cells came of age at the beginning of the century with the use of microarrays to analyze cell populations in bulk. Since then, stem cell transcriptomics has become increasingly sophisticated, notably with the recent widespread use of single-cell RNA sequencing. Here, we provide a perspective on how an early signature of genes upregulated in embryonic and adult stem cells, identified using microarrays over 20 years ago, serendipitously led to the recent discovery that stem/progenitor cells across organs are in a state of hypertranscription, a global elevation of the transcriptome. Looking back, we find that the 2002 stemness signature is a robust marker of stem cell hypertranscription, even though it was developed well before it was known what hypertranscription meant or how to detect it. We anticipate that studies of stem cell hypertranscription will be rich in novel insights in physiological and disease contexts for years to come.

Nucleic acid therapeutics have become increasingly recognized in recent years for their capability to target both coding and non-coding sequences. Several types of nucleic acid modalities, including siRNA, mRNA, aptamer, along with antisense oligo, have been approved by regulatory bodies for therapeutic use. The field of nucleic acid therapeutics has been brought to the forefront by the rapid development of vaccines against COVID-19, followed by a number of approvals for clinical use including much anticipated CRISPR-Cas9. However, obstacles such as the difficulty of achieving efficient and targeted delivery to diseased sites remain. This review provides an overview of nucleic acid therapeutics and highlights substantial advancements, including critical engineering, conjugation, and delivery strategies, that are paving the way for their growing role in modern medicine.

Small interfering RNA (siRNA) drugs were first proposed in 1999. They have reached the market for administration to patients after more than 20 years of development. The US Food and Drug Administration has approved six siRNA drugs in recent years: patisiran, givosiran, lumasiran, vutrisiran, inclisiran, and nedosiran. siRNA drugs are based on the post-transcriptional gene regulation mechanism of RNA interference. These drugs have gained widespread attention for their effectiveness, low dosage, and low frequency of administration. Theoretically, siRNA drugs have great potential due to their ability to silence almost any target gene. However, drug delivery, especially the extrahepatic one, remains a major challenge. Currently, all approved drugs target the liver. The high blood flow, natural filtration function, and drug delivery methods of the liver overall ensure high efficacy and stability of the drugs themselves. This review summarizes the history of siRNA drug development and the mechanisms of action, with a focus on the drug targets, indications, and key clinical trial results to introduce the status of both marketed drugs and those currently in clinical trials. Additionally, this review provides a brief analysis of several key stages of the commercialization process of siRNA drugs.

Chemical modifications are applied to small interfering RNAs (siRNAs) to improve their metabolic stability, specificity, and duration of pharmacodynamic effects. Despite tremendous progress made, identifying chemically modified siRNAs with drug-like properties requires empirical screening due to an intricate interdependence of siRNA sequence and chemistry, i.e., the nature and position of chemical modifications within the siRNA duplex. To improve our ability to design fully modified, potent siRNAs, we combined experimental measurements of thermodynamic stability and biological activity in vitro with extensive molecular modeling in silico of the structural, dynamic, and energetic properties of parent (unmodified) siRNA duplex sequences compared with their chemically modified variants. A pattern of modifications at specific positions were identified, where the combination of sequence and chemical modifications play an outsized role in the observed biological activity. Molecular modeling revealed low stabilization energies and increased sugar stereochemical flexibility for 2'-F modified position g2 and less so for g6 in the guide strand seed region. Machine learning confirmed that these properties correlate with higher observed biological activity. These results provide molecular-level insights into the effects of chemical modifications on the intrinsic activity of siRNAs, which can be used in the rational design of chemically modified siRNAs with uncompromised potency.

Viruses are a major cause of plant disease with an estimated annual economic impact of over $30 billion. They account for nearly 50% of the pathogens responsible for emerging and re-emerging plant diseases worldwide. To confer resistance against these diseases RNA interference (RNAi) technology can be employed. Designing silencing molecules like dsRNA homologous to the viral genome has been the most common non-transgenic method to induce RNAi mediated resistance in plants. dsRNAs are carefully tailored and produced considering factors such as the type of virus, target genomic region, dsRNA size, and method of application to maximise their efficiency. With the advent of new technologies like nano-platforms, a sustainable carrier can be developed to deliver these molecules, enhancing their stability and its bioavailability. This innovative technology faces regulatory debates globally and lacks legislation for commercialisation. But this is an eco-friendly alternative to conventional pesticides that can revolutionize the future of plant viral disease management, providing a bio-safe and an evergreen solution.

The creation of organisms with Cre-loxP conditional gene recombination systems often faces challenges, particularly when creating the initial (F0) generation with both a Cre recombinase and a DNA site flanked by loxP elements (floxed site). The primary reason is that it is difficult to synthesize a single plasmid with both the Cre gene and the floxed site, since Cre-mediated recombination spontaneously occurs when the plasmid is amplified in Escherichia coli bacterial cells. Here, we introduce an artificial nucleic acid sequence TATATATATATATATATA, named TAx9, that enables the integration of both the Cre gene and the floxed site into a single plasmid. TAx9 effectively blocks spontaneous Cre-mediated recombination in E. coli cells. Using this system, we created an F0 generation of transgenic newts and CRISPR-Cas9 knock-in mice with tissue-specific Cre recombination triggered by tamoxifen. TAx9 technology will be a powerful strategy for creating organisms capable of conditional genetic modification in the F0 generation, accelerating various life science research by reducing the time and cost for ultimately establishing and maintaining lines of genetically modified organisms.

Nucleic-acid-based therapies have emerged as a pivotal domain within contemporary biomedical science, marked by significant advancements in recent years. These innovative treatments primarily operate through the precise binding of DNA or RNA molecules to discrete target genes, subsequently suppressing the expression of the target proteins. The spectrum of nucleic-acid-based therapies encompasses antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs), etc. Compared to more traditional medicinal approaches, nucleic-acid-based therapies stand out for their highly targeted action on specific genes, as well as their potential for chemical modification to improve resistance to nucleases, ensuring sustained therapeutic activity and mitigating immunogenicity concerns. Nevertheless, these molecules' limited cellular permeability necessitates the deployment of delivery vectors to enhance their intracellular uptake and stability. As nucleic-acid-based therapies progressively display promising pharmacodynamic profiles, there has been a burgeoning interest in these treatments for applications in clinical research. This review aims to summarize the variety of nucleic acid drugs and their mechanisms, evaluate the present status in research and application, discourse on prospective trends, and potential challenges ahead. These innovative therapeutics are anticipated to assume a pivotal role in the management of a wide array of diseases.

During coevolution, plants produce numerous toxic secondary metabolites to protect themselves. However, most plants still serve as food for insects to meet their nutritional needs. The evolutionary processes that enable herbivorous insects to resist plant defenses remain largely complex and difficult to predict. In this study, lactase phlorizin hydrolase (LPH) was identified for the first time in Aphis gossypii. Bioinformatics analysis showed that the LPH protein belongs to the glycoside hydrolase 1 (GH1) family. The qPCR showed that the transcript level of LPH in cotton aphids treated with the expression dsAgCYP6CY3 cotton lines was significantly reduced by 39.9% at 48 h compared to the nontransgenic cotton, and the hydrolytic activities of LPH on lactose and phlorizin were significantly reduced by 28.7% and 20.1%, respectively. In vitro enzyme activity experiments demonstrated that LPH could hydrolyze lactose and four phenolic glycosides. In addition, RNA interference (RNAi) and insect performance assays showed that silencing LPH affected the growth of the larvae, resulting in the death of the larvae exposed to phenolic glycosides. These results reveal an evolutionary scenario whereby herbivores use deglycosylation to develop resistance to plant defenses and this can be exploited for plant protection.

DOPA decarboxylase (DDC) plays a crucial role in the physiological functions of animals by participating in the dopaminergic system. However, the functions of DDC in shellfish remain poorly understood. The Pacific oyster (Crassostrea gigas) is an extensively cultivated shellfish. In this study, we characterized a DDC gene, designated CgDDC, from C. gigas. The CgDDC gene encodes a protein that contains a Pyridoxal_deC domain, which features specific binding sites for pyridoxal-5'-phosphate (PLP) and L-DOPA. CgDDC exhibits a significantly higher expression level in the black shell oyster strain than the white strain. In vitro enzymatic reaction assays demonstrated that CgDDC catalyzes the conversion of L-DOPA to dopamine. In vivo experiments revealed that inhibiting CgDDC activity reduced the expression of genes associated with tyrosine metabolism. Furthermore, the knockdown of CgDDC caused a decline in cAMP level and reduced transcription of genes involved in the cAMP-mediated melanogenesis. Additionally, treatment with L-α-DOPA inhibited CgDDC enzyme activity and cAMP-mediated melanogenesis; however, dopamine supplementation countered this inhibition, maintaining gene expression and melanin content at baseline levels. Collectively, our findings suggest that CgDDC is intricately involved in regulating tyrosine metabolism and melanogenesis in C. gigas.

Targeted therapy has emerged as a transformative breakthrough in modern medicine. Oligonucleotide drugs, such as antisense oligonucleotides (ASOs) and small interfering RNAs (siRNAs), have made significant advancements in targeted therapy. Other oligonucleotide-based therapeutics like clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) systems are also leading a revolution in targeted gene therapy. However, hybridisation-dependent off-target effects, arising from imperfect base pairing, remain a significant and growing concern for the clinical translation of oligonucleotide-based therapeutics. These mismatches in base pairing can lead to unintended steric blocking or cleavage events in non-pathological genes, affecting the efficacy and safety of the oligonucleotide drugs. In this review, we examine recent developments in oligonucleotide-based targeted therapeutics, explore the factors influencing sequence-dependent targeting specificity, and discuss the current approaches employed to reduce the off-target side effects. The existing strategies, such as chemical modifications and oligonucleotide length optimisation, often require a trade-off between specificity and binding affinity. To further address the challenge of hybridisation-dependent off-target effects, we discuss DNA nanotechnology-based strategies that leverage the collaborative effects of nucleic acid assembly in the design of oligonucleotide-based therapies. In DNA nanotechnology, collaborative effects refer to the cooperative interactions between individual strands or nanostructures, where multiple bindings result in more stable and specific hybridisation behaviour. By requiring multiple complementary interactions to occur simultaneously, the likelihood of unintended partially complementary binding events in nucleic acid hybridisation should be reduced. And thus, with the aid of collaborative effects, DNA nanotechnology has great promise in achieving both high binding affinity and high specificity to minimise the hybridisation-dependent off-target effects of oligonucleotide-based therapeutics.