Nucleic acid therapeutics最新文献

A Workflow for Transcriptome-Wide Assessment of Antisense Oligonucleotide Selectivity. 反义寡核苷酸选择性转录组范围评估工作流程。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-29 DOI: 10.1177/21593337251378141

Sagar S Damle, Andy Watt, Steven Kuntz, Amanda Crutchfield, Emma Carlborg, Judy Webb, Clare Quirk, Dorde Relic, Scott Donovan, Christopher E Hart, Frank Rigo

{"title":"A Workflow for Transcriptome-Wide Assessment of Antisense Oligonucleotide Selectivity.","authors":"Sagar S Damle, Andy Watt, Steven Kuntz, Amanda Crutchfield, Emma Carlborg, Judy Webb, Clare Quirk, Dorde Relic, Scott Donovan, Christopher E Hart, Frank Rigo","doi":"10.1177/21593337251378141","DOIUrl":"https://doi.org/10.1177/21593337251378141","url":null,"abstract":"Antisense oligonucleotides (ASOs) designed to recruit RNase H1 (gapmer ASOs) have been used successfully to downregulate the expression of therapeutic targets. Gapmer ASOs can be identified that selectively reduce the expression of transcripts containing the perfectly complementary intended ASO target site without affecting the expression of unintended transcripts (selective ASOs). However, ASOs can also be identified that reduce the expression of unintended transcripts with target sites that are not perfectly complementary to the ASO (nonselective ASOs). Currently, the understanding of in silico rules for predicting off-targets is suboptimal. In order to determine the selectivity of gapmer ASOs, we therefore developed an experimental workflow called concentration-response digital gene expression (CR-DGE). In CR-DGE, ASO treatment is performed at increasing concentrations, and the effect on the transcriptome is measured using 3'Tag-Seq. Expression data are then analyzed to identify genes with concentration-responsive knockdown. We demonstrate that CR-DGE identifies gapmer ASO concentration-responsive genes with high reproducibility and greater sensitivity than conventional single-concentration assays. Applying CR-DGE to a panel of gapmer ASOs identifies ASOs with a range of selectivity. These results demonstrate that CR-DGE can be used effectively to assess the selectivity of gapmer ASOs, offering a valuable tool for research and therapeutic development.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145186693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cryopreservation of siRNA-Treated Cells Is Feasible. sirna处理细胞的低温保存是可行的。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-23 DOI: 10.1177/21593337251381041

Melanie Sauer, Xavier Segarra-Visent, Leon Breuer, Vasileios Tzirtziganis, Tatyana Ryaykenen, David A Cooper, Dimas Echeverria, Anastasia Kremer, Reka A Haraszti

{"title":"Cryopreservation of siRNA-Treated Cells Is Feasible.","authors":"Melanie Sauer, Xavier Segarra-Visent, Leon Breuer, Vasileios Tzirtziganis, Tatyana Ryaykenen, David A Cooper, Dimas Echeverria, Anastasia Kremer, Reka A Haraszti","doi":"10.1177/21593337251381041","DOIUrl":"https://doi.org/10.1177/21593337251381041","url":null,"abstract":"Cryopreservation is a routine step in the manufacturing process of adoptive cell therapies (ACT), providing critical logistic flexibility. RNA interference (RNAi)-based therapies are increasingly being explored as enhancers or modulators of ACT. However, the impact of cryopreservation on cells treated with RNAi-based therapies has not been investigated before. In this study, we addressed this knowledge gap by examining silencing efficacy in small interfering RNA (siRNA)-treated cells that undergo cryopreservation. Our findings demonstrate that silencing in cryopreserved cells is comparable to that in cells maintained continuously in culture. Moreover, we found that the duration of siRNA exposure plays a significant role in cells that later undergo cryopreservation, with extended exposure improving silencing efficiency. However, this effect diminishes at higher siRNA concentrations. Additionally, we showed that siRNA treatment is feasible at low temperatures (2°C-8°C), and siRNA-treated cells can be cryopreserved for extended periods (at least 1 month) without loss of efficacy. Our work establishes the feasibility of integrating siRNA treatments into current manufacturing processes for ACT.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145131499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantification of mRNA Decay Rates in HeLa and SH-SY5Y Cell Lines Reveals Novel Properties of Membrane Protein Coding Transcripts. HeLa和SH-SY5Y细胞系mRNA衰减率的量化揭示了膜蛋白编码转录物的新特性

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-23 DOI: 10.1177/21593337251377561

Melanie B Martinez, Colton J Williamson, Tareian Cazares, Chunlao Tang, Marjoke F Debets, Pooja Gangras

{"title":"Quantification of mRNA Decay Rates in HeLa and SH-SY5Y Cell Lines Reveals Novel Properties of Membrane Protein Coding Transcripts.","authors":"Melanie B Martinez, Colton J Williamson, Tareian Cazares, Chunlao Tang, Marjoke F Debets, Pooja Gangras","doi":"10.1177/21593337251377561","DOIUrl":"https://doi.org/10.1177/21593337251377561","url":null,"abstract":"Posttranscriptional regulation is crucial for siRNA design, as decay rates in cell lines influence perceived siRNA potency. This study profiles transcripts with 'fast' and 'slow' half-lives in HeLa and SH-SY5Y cells, commonly used in drug discovery. We calculated half-lives for 1,815 HeLa and 5,376 SH-SY5Y transcripts, finding comparable half-lives between cell lines, though HeLa cells generally had longer half-lives. Comparing mRNA and protein half-lives, 'fast' decay transcripts encoded proteins with shorter half-lives, while 'slow' decay transcripts encoded stable proteins. We linked mRNA decay rates to siRNA activity by comparing HeLa data to a previous siRNA screen, discovering that faster decay transcripts had lower knockdown. Surprisingly, stable transcripts, more amenable to knockdown, were over-represented by membrane protein-coding transcripts. Despite their stability, these transcripts had low-to-moderate expression, regardless of miRNA regulation. We explored cis- and trans- features affecting mRNA stability and expression, suggesting that low RNA binding protein (RBP) binding, combined with specific stabilizing RBP regulation, contributes to the stability of these membrane protein-coding transcripts. This study highlights the importance of understanding transcript features, mRNA decay and its potential impact on siRNA efficacy, particularly for transcripts encoding membrane proteins.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145131541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Binding and Ligand Activation Driven Enrichment-Directed Evolution of SaCas9 gRNAs Improves Gene Editing Efficiency. 结合和配体激活驱动的SaCas9 gRNAs富集定向进化提高了基因编辑效率。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-15 DOI: 10.1177/21593337251370553

Telmo Llanga, Korie Bush, Ying Sun, Amy Yan, Jonathan Zhou, Jan Gorodkin, Bruce A Sullenger

{"title":"Binding and Ligand Activation Driven Enrichment-Directed Evolution of SaCas9 gRNAs Improves Gene Editing Efficiency.","authors":"Telmo Llanga, Korie Bush, Ying Sun, Amy Yan, Jonathan Zhou, Jan Gorodkin, Bruce A Sullenger","doi":"10.1177/21593337251370553","DOIUrl":"https://doi.org/10.1177/21593337251370553","url":null,"abstract":"Clustered regularly interspaced short palindromic repeats-based editing is inefficient at over two-thirds of genetic targets. A primary cause is ribonucleic acid (RNA) misfolding that can occur between the spacer and scaffold regions of the gRNA, which hinders the formation of functional Cas9 ribonucleoprotein (RNP) complexes. Here, we uncover hundreds of highly efficient gRNA variant scaffolds for Staphylococcus aureus (Sa)Cas9 utilizing an innovative binding and ligand activation driven enrichment (BLADE) methodology, which leverages asymmetrical product dissociation over rounds of evolution. SaBLADE-derived gRNA scaffolds contain 7%-42% of nucleotide variation relative to wild type. gRNA variants are able to improve gene editing efficiency at all targets tested, and they achieve their highest levels of editing improvement (>400%) at the most challenging DNA target sites for the wild-type SaCas9 gRNA. This arsenal of SaBLADE-derived gRNA variants showcases the power and flexibility of combinatorial chemistry and directed evolution to enable efficient gene editing at challenging, or previously intractable, genomic sites.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145064985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Use of a Transgenic Human PNPLA3^I148M Knock-in Mouse for Translational Safety Evaluations of siRNA Therapeutics. 使用转基因人PNPLA3I148M敲入小鼠进行siRNA疗法的翻译安全性评估。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-06 DOI: 10.1177/21593337251375804

Ben Brooks, Artem Shkumatov, Jackson Kalanzi, Kim Henderson Park, Julie M Lade, Diana Wong, Hui Dou, Jason Long, Ingrid C Rulifson, Justin K Murray, Lauren Mihalcik, Tod A Harper

{"title":"Use of a Transgenic Human PNPLA3I148M Knock-in Mouse for Translational Safety Evaluations of siRNA Therapeutics.","authors":"Ben Brooks, Artem Shkumatov, Jackson Kalanzi, Kim Henderson Park, Julie M Lade, Diana Wong, Hui Dou, Jason Long, Ingrid C Rulifson, Justin K Murray, Lauren Mihalcik, Tod A Harper","doi":"10.1177/21593337251375804","DOIUrl":"10.1177/21593337251375804","url":null,"abstract":"The PNPLA3 single nucleotide polymorphism, rs738409, is the strongest known genetic risk factor for metabolic dysfunction-associated steatotic liver disease; thus, targeting the minor allele with a GalNAc-conjugated siRNA is an attractive strategy to treat patients carrying the genetic variant. To enable translational safety assessment of a GalNAc-conjugated siRNA that specifically targets the rs738409 sequence of PNPLA3, a transgenic human PNPLA3I148M knock-in mouse (huPNPLA3I148M) was utilized. This model showed no significant genotype-related phenotypic differences to wild-type mice in a phenotype characterization study when maintained on standard rodent chow. Additionally, a repeat-dose toxicology study using a GalNAc-conjugated siRNA specific for rs738409 resulted in comparable findings between genotypes (i.e., liver enzyme and histopathology changes), indicating the findings were due to the siRNA therapeutic and not a result of target knockdown in huPNPLA3I148M mice. Overall, these data demonstrate the huPNPLA3I148M mouse is suitable for repeat-dose toxicology studies, suggesting this approach could be applied to other siRNA programs lacking a pharmacologically relevant nonclinical species to support translational safety assessments during drug development.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144963276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cell Type Distribution of Intrathecal Antisense Oligonucleotide Activity in Deep Brain Regions of Non-Human Primates. 非人灵长类动物脑深部鞘内反义寡核苷酸活性的细胞类型分布。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-09-06 DOI: 10.1177/21593337251371594

Jeannine A Frei, Juliana E Gentile, Yuan Lian, Meredith A Mortberg, Juliana Capitanio, Paymaan Jafar-Nejad, Sonia M Vallabh, Hien T Zhao, Eric Vallabh Minikel

{"title":"Cell Type Distribution of Intrathecal Antisense Oligonucleotide Activity in Deep Brain Regions of Non-Human Primates.","authors":"Jeannine A Frei, Juliana E Gentile, Yuan Lian, Meredith A Mortberg, Juliana Capitanio, Paymaan Jafar-Nejad, Sonia M Vallabh, Hien T Zhao, Eric Vallabh Minikel","doi":"10.1177/21593337251371594","DOIUrl":"10.1177/21593337251371594","url":null,"abstract":"Intrathecally administered RNase H1-active gapmer antisense oligonucleotides (ASOs) are promising therapeutics for brain diseases where lowering the expression of one target gene is expected to be therapeutically beneficial. Such ASOs are active, to varying degrees, across most or all cell types in the cortex and cerebellum of mouse and non-human primate (NHP) brain regions with substantial drug accumulation. Intrathecally delivered ASOs, however, exhibit a gradient of exposure across the brain, with more limited drug accumulation and weaker target engagement in deep brain regions of NHP. Here, we profiled the activity of a tool, ASO, against Malat1 in three deep brain regions of NHP: thalamus, caudate, and putamen. All neuronal subtypes exhibited knockdown similar to, or deeper than, the bulk tissue. Among non-neuronal cells, knockdown was deepest in microglia and weakest in endothelial stalk. Overall, we observed broad target engagement across all cell types detected, supporting the relevance of intrathecal ASOs to diseases with deep brain involvement.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144963311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In Vivo Phage Display for the Identification of Muscle Homing Peptides to Improve the Delivery of Phosphorodiamidate Morpholino Oligomers for Duchenne Muscular Dystrophy Therapy. 利用体内噬菌体展示技术鉴定肌肉归一肽，以改善治疗杜氏肌营养不良的磷酸二酯Morpholino低聚物的递送。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-08-29 DOI: 10.1177/21593337251371708

Anne-Fleur E Schneider, Christa L Tanganyika-de Winter, Hailiang Mei, Silvana M G Jirka, Xuyu Tan, Emily G Thompson, Kristin Ha, Anindita Mitra, Stephanie Garcia, Marleen Luimes, Ryan Oliver, Kathy Y Morgan, Vincent Guerlavais, Annemieke Aartsma-Rus

{"title":"In Vivo Phage Display for the Identification of Muscle Homing Peptides to Improve the Delivery of Phosphorodiamidate Morpholino Oligomers for Duchenne Muscular Dystrophy Therapy.","authors":"Anne-Fleur E Schneider, Christa L Tanganyika-de Winter, Hailiang Mei, Silvana M G Jirka, Xuyu Tan, Emily G Thompson, Kristin Ha, Anindita Mitra, Stephanie Garcia, Marleen Luimes, Ryan Oliver, Kathy Y Morgan, Vincent Guerlavais, Annemieke Aartsma-Rus","doi":"10.1177/21593337251371708","DOIUrl":"https://doi.org/10.1177/21593337251371708","url":null,"abstract":"The severe X-linked degenerative neuromuscular disease Duchenne muscular dystrophy (DMD) is caused by the loss of dystrophin through reading frame disruptive mutations in the DMD gene. Dystrophin protein is crucial for the stability of the muscle. Targeting specific exons with antisense oligonucleotides (ASO) will prevent inclusion of the exon during pre-mRNA splicing, which can restore the reading frame, facilitating the production of partially functional dystrophin proteins. For DMD, four ASOs of the phosphorodiamidate morpholino oligomer (PMOs) chemistry are FDA approved. It is anticipated that improved delivery to skeletal muscle and heart will lead to larger therapeutic results. With our research, we sought to identify muscle-homing peptides that can achieve increased delivery of ASOs to muscle or heart when conjugated to PMOs. We applied in vivo phage display biopanning mouse models for DMD to identify muscle-homing peptides while simultaneously negatively selecting peptides that home to unwanted organs, such as the kidney and liver. After confirmation of the muscle homing ability in vitro, we conjugated selected candidate peptides to PMOs to be tested in vivo, where we found that conjugation of one specific muscle homing peptide led to significantly improved delivery to muscle, with a small improvement in exon skipping and dystrophin restoration.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144963327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Context Matters: The Importance of a Comprehensive Genomic Region When Assessing the Therapeutic Potential of Antisense Oligonucleotides in Splicing Assays. 背景问题：在剪接分析中评估反义寡核苷酸的治疗潜力时，综合基因组区域的重要性。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-08-25 DOI: 10.1177/21593337251371582

Dyah W Karjosukarso, Julia F Kiefmann, Femke Bukkems, Lonneke Duijkers, Rob W J Collin

引用次数: 0

Exosome-Mediated Mitochondrial Delivery of Antisense Oligonucleotides. 外泌体介导的反义寡核苷酸的线粒体递送。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-08-01 Epub Date: 2025-06-18 DOI: 10.1089/nat.2024.0067

Dora von Trentini, Ivan J Dmochowski

{"title":"Exosome-Mediated Mitochondrial Delivery of Antisense Oligonucleotides.","authors":"Dora von Trentini, Ivan J Dmochowski","doi":"10.1089/nat.2024.0067","DOIUrl":"10.1089/nat.2024.0067","url":null,"abstract":"We present a general method for in-cellulo delivery of 2'-O-methyl (2'-OMe) RNA oligonucleotides (oligos) to mitochondria for antisense applications, with potential for implementation in other mitochondrial DNA (mtDNA)-targeted therapies. Exosomes, which are nanoscale, naturally occurring extracellular vesicles (EVs), have been employed for biotechnology applications in oligonucleotide delivery in recent years. We discovered that exosomes from fetal bovine serum (FBS) can be used as a simple and biologically compatible delivery agent of 2'-OMe RNA antisense oligonucleotides to cellular mitochondria, leading to target protein knockdown. While most RNA interference and antisense mechanisms occur in the cytoplasm or nucleus, the need for mitochondrial targeting has become increasingly apparent. Mitochondrial disease describes a variety of currently incurable syndromes that especially affect organs requiring significant energy including the muscles, heart, and brain. Many of these syndromes result from mutations in mtDNA, which codes for the 13 proteins of the oxidative phosphorylation system and are thus often implicated in inherited metabolic disorders.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"198-208"},"PeriodicalIF":4.7,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12411094/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144485293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterizing Antibodies Targeting Antisense Oligonucleotide Phosphorothioate and 2'-O-Methoxyethyl Modifications for Intracellular Trafficking and Biodistribution Studies. 针对反义寡核苷酸硫代酸和2'- o -甲氧基乙基修饰的抗体在细胞内运输和生物分布研究中的特征。

IF 4.7 2区医学

Nucleic acid therapeutics Pub Date : 2025-08-01 Epub Date: 2025-07-21 DOI: 10.1177/21593337251361396

Inês Fial, Seth A Farrier, David P Chimento, Carl A Ascoli, Xiao Wan, Peter L Oliver

{"title":"Characterizing Antibodies Targeting Antisense Oligonucleotide Phosphorothioate and 2'-O-Methoxyethyl Modifications for Intracellular Trafficking and Biodistribution Studies.","authors":"Inês Fial, Seth A Farrier, David P Chimento, Carl A Ascoli, Xiao Wan, Peter L Oliver","doi":"10.1177/21593337251361396","DOIUrl":"10.1177/21593337251361396","url":null,"abstract":"The efficacy of nucleic acid therapeutics (NATs) such as antisense oligonucleotides (ASOs) and small interfering RNAs relies on multiple stages of extra- and intracellular trafficking. Assessing uptake and efficacy often relies on fluorescent tagging of the NAT for imaging, although the exogenous tag undoubtedly influences the kinetics of intracellular transport and does not represent the compound used clinically. Therefore, better methods to assess the cellular and tissue distribution of NATs are needed. Here, we have validated new panels of antibody reagents that target clinically relevant nucleic acid modifications for visualizing ASOs both in vitro and in vivo. Using the ModDetect™ library of antibodies, we have tested ASOs in vitro for intracellular localization by immunocytochemistry and for biodistribution in mouse tissues by immunohistochemistry. Antibodies specific for the commonly used phosphorothioate (PS) or 2'-O-methoxyethyl (2'-MOE) modifications successfully detected gapmer ASOs, facilitating colocalization studies with endosomal markers in 2D and 3D cell models. In addition, we assessed colocalization of anti-PS signals with fluorescently tagged ASOs. Our data demonstrate the utility of these reagents for the NAT field, where modified nucleic acids can be detected irrespective of the nucleotide sequence, rendering the system amenable for multiple clinical and preclinical workflows and quantitative immunoassays.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"168-181"},"PeriodicalIF":4.7,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144675391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0