Nucleic acid therapeutics最新文献_第2页

Characterization of the TLR9-Activating Potential of LNA-Modified Antisense Oligonucleotides. LNA修饰的反义寡核苷酸的TLR9激活潜力表征

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-10-01 Epub Date: 2024-07-17 DOI: 10.1089/nat.2024.0013

Irene Riera-Tur, Julia Hinterdobler, André Maaske, Anne Sadewasser, Monika Schell, Janani Sekar, Sven Michel, Richard Klar, Frank Jaschinski

{"title":"Characterization of the TLR9-Activating Potential of LNA-Modified Antisense Oligonucleotides.","authors":"Irene Riera-Tur, Julia Hinterdobler, André Maaske, Anne Sadewasser, Monika Schell, Janani Sekar, Sven Michel, Richard Klar, Frank Jaschinski","doi":"10.1089/nat.2024.0013","DOIUrl":"10.1089/nat.2024.0013","url":null,"abstract":"Early characterization of the immunostimulatory potential of therapeutic antisense oligonucleotides (ASOs) is crucial. At present, little is known about the toll-like receptor 9 (TLR9)-mediated immunostimulatory potential of third-generation locked nucleic acid (LNA)-modified ASOs. In this study, we have systematically investigated the TLR9-activating potential of LNA-modified oligonucleotides using different mouse and human cell culture systems. Although it has been reported that LNA modifications as well as cytosine methylation of 5'-cytosine-phosphate-guanine-3' (CpG) motifs can reduce TLR9 stimulation by phosphorothioate (PTO)-modified oligonucleotides, we identified CpG-containing LNA gapmers with substantial TLR9-stimulatory activity. We further identified immunostimulatory LNA gapmers without CpG motifs. Unexpectedly, methylation of cytosines only within the CpG motif did not necessarily reduce but could even increase TLR9 activation. In contrast, systematic methylation of all cytosines reduced or even abrogated TLR9 activation in most cases. Context dependently, the introduction of LNA-modifications into the flanks could either increase or decrease TLR9 stimulation. Overall, our results indicate that TLR9-dependent immunostimulatory potential is an individual feature of an oligonucleotide and needs to be investigated on a case-by-case basis.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"257-271"},"PeriodicalIF":4.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141634130","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

Antisense Oligonucleotide STK-002 Increases OPA1 in Retina and Improves Mitochondrial Function in Autosomal Dominant Optic Atrophy Cells. 反义寡核苷酸 STK-002 增加视网膜中的 OPA1 并改善常染色体显性视神经萎缩细胞的线粒体功能

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-09-12 DOI: 10.1089/nat.2024.0022

Aditya Venkatesh,Taylor McKenty,Syed Ali,Donna Sonntag,Shobha Ravipaty,Yanyan Cui,Deirdre Slate,Qian Lin,Anne Christiansen,Sarah Jacobson,Jacob Kach,Kian Huat Lim,Vaishnavi Srinivasan,Boris Zinshteyn,Isabel Aznarez,Laryssa A Huryn,Zhiyu Li,Robert B Hufnagel,Gene Liau,Karen Anderson,Jeff Hoger

{"title":"Antisense Oligonucleotide STK-002 Increases OPA1 in Retina and Improves Mitochondrial Function in Autosomal Dominant Optic Atrophy Cells.","authors":"Aditya Venkatesh,Taylor McKenty,Syed Ali,Donna Sonntag,Shobha Ravipaty,Yanyan Cui,Deirdre Slate,Qian Lin,Anne Christiansen,Sarah Jacobson,Jacob Kach,Kian Huat Lim,Vaishnavi Srinivasan,Boris Zinshteyn,Isabel Aznarez,Laryssa A Huryn,Zhiyu Li,Robert B Hufnagel,Gene Liau,Karen Anderson,Jeff Hoger","doi":"10.1089/nat.2024.0022","DOIUrl":"https://doi.org/10.1089/nat.2024.0022","url":null,"abstract":"Autosomal dominant optic atrophy (ADOA) is an inherited optic neuropathy most frequently associated with OPA1 mutations. Most variants result in haploinsufficiency, and patient cells express roughly half of the normal levels of OPA1 protein. OPA1 is a mitochondrial GTPase that is essential for normal mitochondrial function. We identified and characterized STK-002, an antisense oligonucleotide (ASO) designed to prevent the incorporation of a naturally occurring alternatively spliced nonproductive exon in OPA1. STK-002 dose dependently reduced the inclusion of this exon, and increased OPA1 protein in human cells, including ADOA patient-derived fibroblasts. ADOA patient cells manifest reduced mitochondrial respiration, and treatment with STK-002 improved the parameters of mitochondrial respiratory function in these cells. Since STK-002 increases OPA1 through the wild-type allele, we assessed retinal OPA1 in wild-type cynomolgus monkeys and rabbits after intravitreal administration of STK-002 or a rabbit-specific surrogate. Increased OPA1 protein was produced in retinal tissue in both species at 4 weeks after ASO injection and persisted in monkeys at 8 weeks. STK-002 and enhanced OPA1 immunofluorescence were visualized in retinal ganglion cells of cynomolgus monkeys treated with the ASO. Cumulatively, these data support the progression of STK-002 toward the clinic as the first potential disease-modifying treatment for ADOA.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"44 1","pages":""},"PeriodicalIF":4.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142263110","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

Screening Splice-Switching Antisense Oligonucleotides in Pancreas-Cancer Organoids. 在胰腺癌组织细胞中筛选剪接转换反义寡核苷酸

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-08-01 Epub Date: 2024-05-08 DOI: 10.1089/nat.2023.0070

Ledong Wan, Alexander J Kral, Dillon Voss, Balázs Schäfer, Kavitha Sudheendran, Mathias Danielsen, Marvin H Caruthers, Adrian R Krainer

{"title":"Screening Splice-Switching Antisense Oligonucleotides in Pancreas-Cancer Organoids.","authors":"Ledong Wan, Alexander J Kral, Dillon Voss, Balázs Schäfer, Kavitha Sudheendran, Mathias Danielsen, Marvin H Caruthers, Adrian R Krainer","doi":"10.1089/nat.2023.0070","DOIUrl":"10.1089/nat.2023.0070","url":null,"abstract":"Aberrant alternative splicing is emerging as a cancer hallmark and a potential therapeutic target. It is the result of dysregulated or mutated splicing factors, or genetic alterations in splicing-regulatory cis-elements. Targeting individual altered splicing events associated with cancer-cell dependencies is a potential therapeutic strategy, but several technical limitations need to be addressed. Patient-derived organoids are a promising platform to recapitulate key aspects of disease states, and to facilitate drug development for precision medicine. Here, we report an efficient antisense-oligonucleotide (ASO) lipofection method to systematically evaluate and screen individual splicing events as therapeutic targets in pancreatic ductal adenocarcinoma organoids. This optimized delivery method allows fast and efficient screening of ASOs, e.g., those that reverse oncogenic alternative splicing. In combination with advances in chemical modifications of oligonucleotides and ASO-delivery strategies, this method has the potential to accelerate the discovery of antitumor ASO drugs that target pathological alternative splicing.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"188-198"},"PeriodicalIF":4.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11387002/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140876860","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

Preclinical Pharmacokinetics in Tumors and Normal Tissues of the Antigene PNA Oligonucleotide MYCN-Inhibitor BGA002. 抗原 PNA 寡核苷酸 MYCN 抑制剂 BGA002 在肿瘤和正常组织中的临床前药代动力学。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-08-01 Epub Date: 2024-07-03 DOI: 10.1089/nat.2024.0005

Anna Lisa Scardovi, Damiano Bartolucci, Luca Montemurro, Sonia Bortolotti, Silvia Angelucci, Camilla Amadesi, Giammario Nieddu, Sean Oosterholt, Lucia Cerisoli, Oscar Della Pasqua, Patrizia Hrelia, Roberto Tonelli

{"title":"Preclinical Pharmacokinetics in Tumors and Normal Tissues of the Antigene PNA Oligonucleotide MYCN-Inhibitor BGA002.","authors":"Anna Lisa Scardovi, Damiano Bartolucci, Luca Montemurro, Sonia Bortolotti, Silvia Angelucci, Camilla Amadesi, Giammario Nieddu, Sean Oosterholt, Lucia Cerisoli, Oscar Della Pasqua, Patrizia Hrelia, Roberto Tonelli","doi":"10.1089/nat.2024.0005","DOIUrl":"10.1089/nat.2024.0005","url":null,"abstract":"Although MYCN has been considered an undruggable target, MYCN alterations confer poor prognosis in many pediatric and adult cancers. The novel MYCN-speciﬁc inhibitor BGA002 is an antigene peptide nucleic acid oligonucleotide covalently bound to a nuclear localization signal peptide. In the present study, we characterized the pharmacokinetics (PK) of BGA002 after single and repeated administration to mice using a novel specific enzyme-linked immunosorbent assay. BGA002 concentrations in plasma showed linear PK, with dose proportional increase across the tested dose levels and similar exposure between male and female and between intravenous and subcutaneous route of administration. Repeated dosing resulted in no accumulation in plasma. Biodistribution up to 7 days after single subcutaneous administration of [14C]-radiolabeled BGA002 showed broad tissues and organ distribution (suggesting a potential capability to reach primary tumor and metastasis in several body sites), with high concentrations in kidney, liver, spleen, lymph nodes, adrenals, and bone marrow. Remarkably, we demonstrated that BGA002 concentrates in tumors after repeated systemic administrations in three mouse models with MYCN amplification (neuroblastoma, rhabdomyosarcoma, and small-cell lung cancer), leading to a significant reduction in tumor weight. Taking into account the available safety profile of BGA002, these data support further evaluation of BGA002 in patients with MYCN-positive tumors.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"173-187"},"PeriodicalIF":4.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141492887","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

mRNA Nuclear Clustering Leads to a Difference in Mutant Huntingtin mRNA and Protein Silencing by siRNAs In Vivo. mRNA 核集群导致体内 siRNAs 沉默突变型亨廷汀 mRNA 和蛋白的差异。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-08-01 Epub Date: 2024-07-18 DOI: 10.1089/nat.2024.0027

Sarah Allen, Daniel O'Reilly, Rachael Miller, Ellen Sapp, Ashley Summers, Joseph Paquette, Dimas Echeverria Moreno, Brianna Bramato, Nicholas McHugh, Ken Yamada, Neil Aronin, Marian DiFiglia, Anastasia Khvorova

{"title":"mRNA Nuclear Clustering Leads to a Difference in Mutant Huntingtin mRNA and Protein Silencing by siRNAs In Vivo.","authors":"Sarah Allen, Daniel O'Reilly, Rachael Miller, Ellen Sapp, Ashley Summers, Joseph Paquette, Dimas Echeverria Moreno, Brianna Bramato, Nicholas McHugh, Ken Yamada, Neil Aronin, Marian DiFiglia, Anastasia Khvorova","doi":"10.1089/nat.2024.0027","DOIUrl":"10.1089/nat.2024.0027","url":null,"abstract":"Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by CAG repeat expansion in the first exon of the huntingtin gene (HTT). Oligonucleotide therapeutics, such as short interfering RNA (siRNA), reduce levels of huntingtin mRNA and protein in vivo and are considered a viable therapeutic strategy. However, the extent to which they silence huntingtin mRNA in the nucleus is not established. We synthesized siRNA cross-reactive to mouse (wild-type) Htt and human (mutant) HTT in a divalent scaffold and delivered to two mouse models of HD. In both models, divalent siRNA sustained lowering of wild-type Htt, but not mutant HTT mRNA expression in striatum and cortex. Near-complete silencing of both mutant HTT protein and wild-type HTT protein was observed in both models. Subsequent fluorescent in situ hybridization analysis shows that divalent siRNA acts predominantly on cytoplasmic mutant HTT transcripts, leaving clustered mutant HTT transcripts in the nucleus largely intact in treated HD mouse brains. The observed differences between mRNA and protein levels, exaggerated in the case of extended repeats, might apply to other repeat-associated neurological disorders.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":" ","pages":"164-172"},"PeriodicalIF":4.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11387003/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141634131","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

Phosphorothioates and Me: A Lecture About My 35 Years in Oligo-World on My Receipt of the 2023 Lifetime Achievement Award of the Oligonucleotide Therapeutics Society. 我与硫代磷酸酯寡核苷酸治疗学会 2023 年终身成就奖获得者关于我在寡核苷酸世界 35 年的演讲。

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-08-01 Epub Date: 2024-06-26 DOI: 10.1089/nat.2024.0032

Cy A Stein

引用次数: 0

Sequence- and Structure-Dependent Cytotoxicity of Phosphorothioate and 2'-O-Methyl Modified Single-Stranded Oligonucleotides. 硫代磷酸酯和 2'-O- 甲基修饰单链寡核苷酸的序列和结构依赖性细胞毒性

IF 4 2区医学

Nucleic acid therapeutics Pub Date : 2024-04-22 DOI: 10.1089/nat.2023.0056

L. Croft, Mark Fisher, Tabassum Khair Barbhuiya, Serene El-Kamand, Samuel Beard, Aleksandra Rajapakse, Roland Gamsjaeger, L. Cubeddu, E. Bolderson, Ken O’Byrne, Derek Richard, Neha S Gandhi

{"title":"Sequence- and Structure-Dependent Cytotoxicity of Phosphorothioate and 2'-O-Methyl Modified Single-Stranded Oligonucleotides.","authors":"L. Croft, Mark Fisher, Tabassum Khair Barbhuiya, Serene El-Kamand, Samuel Beard, Aleksandra Rajapakse, Roland Gamsjaeger, L. Cubeddu, E. Bolderson, Ken O’Byrne, Derek Richard, Neha S Gandhi","doi":"10.1089/nat.2023.0056","DOIUrl":"https://doi.org/10.1089/nat.2023.0056","url":null,"abstract":"Single-stranded oligonucleotides (SSOs) are a rapidly expanding class of therapeutics that comprises antisense oligonucleotides, microRNAs, and aptamers, with ten clinically approved molecules. Chemical modifications such as the phosphorothioate backbone and the 2'-O-methyl ribose can improve the stability and pharmacokinetic properties of therapeutic SSOs, but they can also lead to toxicity in vitro and in vivo through nonspecific interactions with cellular proteins, gene expression changes, disturbed RNA processing, and changes in nuclear structures and protein distribution. In this study, we screened a mini library of 277 phosphorothioate and 2'-O-methyl-modified SSOs, with or without mRNA complementarity, for cytotoxic properties in two cancer cell lines. Using circular dichroism, nucleic magnetic resonance, and molecular dynamics simulations, we show that phosphorothioate- and 2'-O-methyl-modified SSOs that form stable hairpin structures through Watson-Crick base pairing are more likely to be cytotoxic than those that exist in an extended conformation. In addition, moderate and highly cytotoxic SSOs in our dataset have a higher mean purine composition than pyrimidine. Overall, our study demonstrates a structure-cytotoxicity relationship and indicates that the formation of stable hairpins should be a consideration when designing SSOs toward optimal therapeutic profiles.","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"15 1","pages":""},"PeriodicalIF":4.0,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140674439","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

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