Nucleic acid therapeutics最新文献_第4页

Therapeutic siRNA Loaded to RISC as Single and Double Strands Requires an Appropriate Quantitative Assay for RISC PK Assessment. 以单链和双链形式载入 RISC 的治疗 siRNA 需要适当的定量检测方法来进行 RISC PK 评估。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-19 DOI: 10.1089/nat.2023.0067

Rui Xu, Emmanuel Njumbe Ediage, Tom Verhaeghe, Jan Snoeys, Lieve Dillen

{"title":"Therapeutic siRNA Loaded to RISC as Single and Double Strands Requires an Appropriate Quantitative Assay for RISC PK Assessment.","authors":"Rui Xu, Emmanuel Njumbe Ediage, Tom Verhaeghe, Jan Snoeys, Lieve Dillen","doi":"10.1089/nat.2023.0067","DOIUrl":"https://doi.org/10.1089/nat.2023.0067","url":null,"abstract":"In recent years, therapeutic siRNA projects are booming in the biotech and pharmaceutical industries. As these drugs act by silencing the target gene expression, a critical step is the binding of antisense strands of siRNA to RNA-induced silencing complex (RISC) and then degrading their target mRNA. However, data that we recently obtained suggest that double-stranded siRNA can also load to RISC. This brings a new understanding of the mechanism of RISC loading which may have a potential impact on how quantification of RISC loaded siRNA should be performed. By combining RNA immune precipitation and probe-based hybridization LC-fluorescence approach, we have developed a novel assay that can accurately quantify the RISC-bound antisense strand, irrespective of which form (double-stranded or single-stranded) is loaded on RISC. In addition, this novel assay can discriminate between the 5'-phosphorylated antisense (5'p-AS) and the nonphosphorylated forms, therefore specifically quantifying the RISC bound 5'p-AS. In comparison, stem-loop qPCR assay does not provide discrimination and accurate quantification when the oligonucleotide analyte exists as a mixture of double and single-stranded forms. Taking together, RISC loading assay with probe-hybridization LC-fluorescence technique would be a more accurate and specific quantitative approach for RISC-associated pharmacokinetic assessment.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" 59","pages":""},"PeriodicalIF":4.0,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140683834","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

Correction to: Understanding and Rescuing the Splicing Defect Caused by the Frequent ABCA4 Variant c.4253 + 43G>A Underlying Stargardt Disease, by Nuria Suárez-Herrera et al., Nucleic Acid Ther 2024;34(2):73-82; doi: 10.1089/nat.2023.0076. 更正：理解并挽救由频繁出现的 ABCA4 变异 c.4253 + 43G>A 导致的剪接缺陷（Stargardt 病的基础），作者 Nuria Suárez-Herrera 等，《核酸治疗学》2024;34(2):73-82; doi: 10.1089/nat.2023.0076。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-17 DOI: 10.1089/nat.2023.0076.correx

引用次数: 0

Splice-Switching Antisense Oligonucleotides Correct Phenylalanine Hydroxylase Exon 11 Skipping Defects and Rescue Enzyme Activity in Phenylketonuria. 剪接转换反义寡核苷酸可纠正苯丙氨酸羟化酶外显子 11 跳越缺陷并恢复苯丙酮尿症的酶活性。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-09 DOI: 10.1089/nat.2024.0014

A. Martínez-Pizarro, Mar Alvarez, M. Dembic, Caroline A Lindegaard, Margarita Castro, Eva Richard, Brage S Andresen, L. Desviat

{"title":"Splice-Switching Antisense Oligonucleotides Correct Phenylalanine Hydroxylase Exon 11 Skipping Defects and Rescue Enzyme Activity in Phenylketonuria.","authors":"A. Martínez-Pizarro, Mar Alvarez, M. Dembic, Caroline A Lindegaard, Margarita Castro, Eva Richard, Brage S Andresen, L. Desviat","doi":"10.1089/nat.2024.0014","DOIUrl":"https://doi.org/10.1089/nat.2024.0014","url":null,"abstract":"The PAH gene encodes the hepatic enzyme phenylalanine hydroxylase (PAH), and its deficiency, known as phenylketonuria (PKU), leads to neurotoxic high levels of phenylalanine. PAH exon 11 is weakly defined, and several missense and intronic variants identified in patients affect the splicing process. Recently, we identified a novel intron 11 splicing regulatory element where U1snRNP binds, participating in exon 11 definition. In this work, we describe the implementation of an antisense strategy targeting intron 11 sequences to correct the effect of PAH mis-splicing variants. We used an in vitro assay with minigenes and identified splice-switching antisense oligonucleotides (SSOs) that correct the exon skipping defect of PAH variants c.1199+17G>A, c.1199+20G>C, c.1144T>C, and c.1066-3C>T. To examine the functional rescue induced by the SSOs, we generated a hepatoma cell model with variant c.1199+17G>A using CRISPR/Cas9. The edited cell line reproduces the exon 11 skipping pattern observed from minigenes, leading to reduced PAH protein levels and activity. SSO transfection results in an increase in exon 11 inclusion and corrects PAH deficiency. Our results provide proof of concept of the potential therapeutic use of a single SSO for different exonic and intronic splicing variants causing PAH exon 11 skipping in PKU.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"62 1","pages":""},"PeriodicalIF":4.0,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140723863","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

Considerations for Creating the Next Generation of RNA Therapeutics: Oligonucleotide Chemistry and Innate Immune Responses to Nucleic Acids. 创造下一代 RNA 疗法的考虑因素：寡核苷酸化学与核酸的先天免疫反应。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-01 DOI: 10.1089/nat.2024.29009.sud

Sudhir Agrawal

引用次数: 0

Understanding and Rescuing the Splicing Defect Caused by the Frequent ABCA4 Variant c.4253+43G>A Underlying Stargardt Disease. 了解并挽救由频发 ABCA4 变异 c.4253+43G>A 导致的剪接缺陷，它是斯塔加特病的基础。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-01 Epub Date: 2024-03-12 DOI: 10.1089/nat.2023.0076

Nuria Suárez-Herrera, Alejandro Garanto, Rob W J Collin

{"title":"Understanding and Rescuing the Splicing Defect Caused by the Frequent ABCA4 Variant c.4253+43G>A Underlying Stargardt Disease.","authors":"Nuria Suárez-Herrera, Alejandro Garanto, Rob W J Collin","doi":"10.1089/nat.2023.0076","DOIUrl":"10.1089/nat.2023.0076","url":null,"abstract":"Pathogenic variants in ABCA4 are the underlying molecular cause of Stargardt disease (STGD1), an autosomal recessive macular dystrophy characterized by a progressive loss of central vision. Among intronic ABCA4 variants, c.4253+43G>A is frequently detected in STGD1 cases and is classified as a hypomorphic allele, generally associated with late-onset cases. This variant was previously reported to alter splicing regulatory sequences, but the splicing outcome is not fully understood yet. In this study, we attempted to better understand its effect on splicing and to rescue the aberrant splicing via antisense oligonucleotides (AONs). Wild-type and c.4253+43G>A variant-harboring maxigene vectors revealed additional skipping events, which were not previously detected upon transfection in HEK293T cells. To restore exon inclusion, we designed a set of 27 AONs targeting either splicing silencer motifs or the variant region and screened these in maxigene-transfected HEK293T cells. Candidate AONs able to promote exon inclusion were selected for further testing in patient-derived photoreceptor precursor cells. Surprisingly, no robust splicing modulation was observed in this model system. Overall, this research helped to adequately characterize the splicing alteration caused by the c.4253+43G>A variant, although future development of AON-mediated exon inclusion therapy for ABCA4 is needed.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"73-82"},"PeriodicalIF":4.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140102082","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

Simplified Oligonucleotide Phosphorus Deprotection Process with Reduced 3-(2-Cyanoethyl) Thymidine Impurities. 减少 3-(2-氰乙基)胸苷杂质的简化寡核苷酸去磷工艺。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-01 Epub Date: 2024-02-05 DOI: 10.1089/nat.2023.0060

Xuan Zhou, Xianglin Shi, Yannick Fillon, Firoz Antia, Thomas Pickel, Jing Yang, William Zhang, Armin Delavari, Jiabao Zhang

引用次数: 0

Nucleic Acid Therapeutics: Successes, Milestones, and Upcoming Innovation. 核酸疗法：成功、里程碑和即将到来的创新。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-01 Epub Date: 2024-03-20 DOI: 10.1089/nat.2023.0068

Jillian Belgrad, Hassan H Fakih, Anastasia Khvorova

引用次数: 0

RNA Interference Effectors Selectively Silence the Pathogenic Variant GNAO1 c.607 G > A In Vitro. RNA 干扰效应器可选择性地抑制体外致病变体 GNAO1 c.607 G > A。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-01 Epub Date: 2024-01-12 DOI: 10.1089/nat.2023.0043

Natalia V Klementieva, Evgenii A Lunev, Anna A Shmidt, Elizaveta M Loseva, Irina M Savchenko, Ekaterina A Svetlova, Ivan I Galkin, Anna V Polikarpova, Evgeny V Usachev, Svetlana G Vassilieva, Valeria I Marina, Marina A Dzhenkova, Anna D Romanova, Anton V Agutin, Anna A Timakova, Denis A Reshetov, Tatiana V Egorova, Maryana V Bardina

{"title":"RNA Interference Effectors Selectively Silence the Pathogenic Variant GNAO1 c.607 G > A In Vitro.","authors":"Natalia V Klementieva, Evgenii A Lunev, Anna A Shmidt, Elizaveta M Loseva, Irina M Savchenko, Ekaterina A Svetlova, Ivan I Galkin, Anna V Polikarpova, Evgeny V Usachev, Svetlana G Vassilieva, Valeria I Marina, Marina A Dzhenkova, Anna D Romanova, Anton V Agutin, Anna A Timakova, Denis A Reshetov, Tatiana V Egorova, Maryana V Bardina","doi":"10.1089/nat.2023.0043","DOIUrl":"10.1089/nat.2023.0043","url":null,"abstract":"RNA interference (RNAi)-based therapeutics hold the potential for dominant genetic disorders, enabling sequence-specific inhibition of pathogenic gene products. We aimed to direct RNAi for the selective suppression of the heterozygous GNAO1 c.607 G > A variant causing GNAO1 encephalopathy. By screening short interfering RNA (siRNA), we showed that GNAO1 c.607G>A is a druggable target for RNAi. The si1488 candidate achieved at least twofold allelic discrimination and downregulated mutant protein to 35%. We created vectorized RNAi by incorporating the si1488 sequence into the short hairpin RNA (shRNA) in the adeno-associated virus (AAV) vector. The shRNA stem and loop were modified to improve the transcription, processing, and guide strand selection. All tested shRNA constructs demonstrated selectivity toward mutant GNAO1, while tweaking hairpin structure only marginally affected the silencing efficiency. The selectivity of shRNA-mediated silencing was confirmed in the context of AAV vector transduction. To conclude, RNAi effectors ranging from siRNA to AAV-RNAi achieve suppression of the pathogenic GNAO1 c.607G>A and discriminate alleles by the single-nucleotide substitution. For gene therapy development, it is crucial to demonstrate the benefit of these RNAi effectors in patient-specific neurons and animal models of the GNAO1 encephalopathy.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"90-99"},"PeriodicalIF":4.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139432811","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

"Goldilocks Modifications" for mRNA Therapeutics Won the Nobel Prize. 用于 mRNA 治疗的 "金发修饰 "荣获诺贝尔奖。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-02-01 Epub Date: 2024-01-30 DOI: 10.1089/nat.2023.0062

Li Li

引用次数: 0

Report of the European Medicines Agency Conference on RNA-Based Medicines. 欧洲药品管理局关于基于 RNA 的药物会议的报告。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-02-01 Epub Date: 2024-01-04 DOI: 10.1089/nat.2023.0021

Falk Ehmann, Andreas Kuhn, Anna Maria Gerdina Pasmooij, Anthony Humphreys, Arjon Van Hengel, Brian Dooley, Brigitte Anliker, Camilla Svensson, Daniel Capaldi, David Henshall, Emer Cooke, Haiyan Zhou, Hilde Bastaerts, Jeske Smink, Joop Van Gerven, Leonor Enes, Lubomir Nechev, Marcel Hoefnagel, Mariëtte Driessens, Michael Wenger, Oriane Blanquie, Pawel Widomski, Ralf Herold, René Thürmer, Sol Ruiz, Steffen Thirstrup, Susan Goody, Tal Zaks, Valentina Cordò, Annemieke M Aartsma-Rus

{"title":"Report of the European Medicines Agency Conference on RNA-Based Medicines.","authors":"Falk Ehmann, Andreas Kuhn, Anna Maria Gerdina Pasmooij, Anthony Humphreys, Arjon Van Hengel, Brian Dooley, Brigitte Anliker, Camilla Svensson, Daniel Capaldi, David Henshall, Emer Cooke, Haiyan Zhou, Hilde Bastaerts, Jeske Smink, Joop Van Gerven, Leonor Enes, Lubomir Nechev, Marcel Hoefnagel, Mariëtte Driessens, Michael Wenger, Oriane Blanquie, Pawel Widomski, Ralf Herold, René Thürmer, Sol Ruiz, Steffen Thirstrup, Susan Goody, Tal Zaks, Valentina Cordò, Annemieke M Aartsma-Rus","doi":"10.1089/nat.2023.0021","DOIUrl":"10.1089/nat.2023.0021","url":null,"abstract":"RNA-based medicines have potential to treat a large variety of diseases, and research in the field is very dynamic. Proactively, The European Medicines Agency (EMA) organized a virtual conference on February 2, 2023 to promote the development of RNA-based medicines. The initiative addresses the goal of the EMA Regulatory Science Strategy to 2025 to \"catalyse the integration of science and technology in medicines development.\" The conference focused on RNA technologies (excluding RNA vaccines) and involved different stakeholders, including representatives from academia, industry, regulatory authorities, and patient organizations. The conference comprised presentations and discussion sessions conducted by panels of subject matter experts. In this meeting report, we summarize the presentations and recap the main themes of the panel discussions.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"4-11"},"PeriodicalIF":4.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139087866","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