IF 5 3区生物学

RNA Pub Date : 2025-09-17 DOI: 10.1261/rna.080644.125

Nikoleta Giarimoglou, Adamantia Kouvela, Athanasios Papakyriakou, Jinwei Zhang, Vassiliki Stamatopoulou, Constantinos Stathopoulos

{"title":"Specific targeting of transcriptional T-box riboswitches leads to effective inhibition of S. aureus.","authors":"Nikoleta Giarimoglou, Adamantia Kouvela, Athanasios Papakyriakou, Jinwei Zhang, Vassiliki Stamatopoulou, Constantinos Stathopoulos","doi":"10.1261/rna.080644.125","DOIUrl":"https://doi.org/10.1261/rna.080644.125","url":null,"abstract":"T-box riboswitches belong to a specific class of RNA regulatory elements that control gene expression in Gram-positive bacteria, including prominent human pathogens. They sense the availability of amino acids by detecting the aminoacylation status of their cognate tRNAs and regulate the expression of genes involved in aminoacylation, amino acid transport, and metabolism. Recent advances on the structures and mechanisms of several regulatory non-coding RNAs among pathogenic bacteria have garnered attention for the development of a new generation of species-specific antibacterials. The frequently acquired resistance against current antibiotics has emerged as a significant challenge for healthcare systems and a serious threat to public health. Herein, we report the characterization of an effective T-box riboswitch inhibitor, termed T-box-i, which efficiently disrupts T-box riboswitch-mediated transcription in vivo. T-box-i was selected through a virtual screening campaign of commercially available small molecules against high-resolution crystallographic structures of T-box riboswitches. It exhibited no cytotoxicity in mammalian cells nor induced antibiotic resistance in S. aureus cultures. These findings provide valuable insights into exploiting T-box riboswitches as antibiotic targets and underscore the therapeutic potential of compounds that selectively target extensively structured regulatory RNA elements and interfaces to combat drug-resistant pathogens.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":""},"PeriodicalIF":5.0,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145081498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Polyadenylation landscape of in vivo long-term potentiation in the rat brain. 大鼠脑内多聚腺苷化景观的长期增强。

IF 5 3区生物学

RNA Pub Date : 2025-09-17 DOI: 10.1261/rna.080485.125

Natalia Gumińska, Francois P Pauzin, Bożena Kuźniewska, Jacek Miłek, Patrycja Wardaszka-Pianka, Paweł S Krawczyk, Seweryn Mroczek, Sebastian Jeleń, Patrick U Pagenhart, Clive R Bramham, Andrzej Dziembowski, Magdalena Dziembowska

{"title":"Polyadenylation landscape of in vivo long-term potentiation in the rat brain.","authors":"Natalia Gumińska, Francois P Pauzin, Bożena Kuźniewska, Jacek Miłek, Patrycja Wardaszka-Pianka, Paweł S Krawczyk, Seweryn Mroczek, Sebastian Jeleń, Patrick U Pagenhart, Clive R Bramham, Andrzej Dziembowski, Magdalena Dziembowska","doi":"10.1261/rna.080485.125","DOIUrl":"https://doi.org/10.1261/rna.080485.125","url":null,"abstract":"Local protein synthesis in neurons is vital for synaptic maintenance and plasticity, yet the regulatory mechanisms, particularly cytoplasmic polyadenylation, are not fully understood. This study employed nanopore sequencing to examine transcriptomic responses and 3'-end dynamics in rat hippocampal long-term potentiation (LTP) in vivo and in synaptoneurosomes after in vitro stimulation. Our long-read transcriptomic dataset allows for detailed analysis of mRNA 3'-ends, poly(A) tail lengths, and nucleotide composition. We observed dynamic shifts in polyadenylation site preference post-LTP induction, with significant poly(A) tail lengthening restricted to transcriptionally induced mRNAs. The poly(A) tails of these genes showed increased non-adenosine abundance. In synaptoneurosomes, chemical stimulation led to the shortening of poly(A) tails on preexisting mRNAs, indicating translation-induced deadenylation. This also includes transcripts, which were previously reported to undergo stimulation-induced cytoplasmic polyadenylation, like Camk2a. Additionally, we discovered a group of neuronal transcripts with poly(A) tails abundant in non-adenosine residues. These tails are semi-templated and derived from extremely adenosine-rich 3'UTRs. This study provides a comprehensive overview of mRNA 3'-end dynamics during LTP, offering insights into post-transcriptional regulation following synaptic activation of plasticity in neurons.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":""},"PeriodicalIF":5.0,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145081494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bacterial/archaeal protein-only RNase P: complementation in Escherichia coli uncovers coevolution of protein-only RNase P and precursor tRNA structures in Aquifex aeolicus. 细菌/古细菌蛋白-纯RNase P：在大肠杆菌中的互补揭示了水蚤中蛋白-纯RNase P和前体tRNA结构的共同进化。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080492.125

Swetlana Davydov, Nadine B Wäber, Markus Gößringer, Paul Klemm, Isabell Rennar, Marcus Lechner, Roland K Hartmann

{"title":"Bacterial/archaeal protein-only RNase P: complementation in Escherichia coli uncovers coevolution of protein-only RNase P and precursor tRNA structures in Aquifex aeolicus.","authors":"Swetlana Davydov, Nadine B Wäber, Markus Gößringer, Paul Klemm, Isabell Rennar, Marcus Lechner, Roland K Hartmann","doi":"10.1261/rna.080492.125","DOIUrl":"10.1261/rna.080492.125","url":null,"abstract":"The family of RNase P endoribonucleases comprises diverse enzyme architectures ranging from complex ribonucleoprotein assemblies to single polypeptides as small as ∼23 kDa termed homologs of Aquifex RNase P (HARPs). The HARPs of two hyperthermophilic bacteria (Aquifex aeolicus and Thermodesulfatator indicus) and one thermophilic archaeon (Methanothermobacter thermautotrophicus) restore (although at reduced growth rate) the viability of Escherichia coli cells with a lethal knockdown of its endogenous RNA-based RNase P. Potential causes for retarded growth were analyzed by RNA-seq, northern blot, and primer extension. This revealed inefficient processing by HARPs in the E. coli host, particularly for precursor tRNAs (pre-tRNAs) with acceptor stems extended by one or more G-C base pairs, and also for the non-tRNA substrate pre-4.5S RNA. Yet, E. coli pre-tRNAs fortuitously carrying an A residue immediately upstream of the (canonical or aberrant) cleavage site were processed more efficiently, particularly by the two bacterial HARPs. Follow-up in vitro processing assays using RNase P model substrates confirmed that an A residue immediately upstream of the cleavage site increases efficiency and accuracy in reactions catalyzed by HARPs. In the case of A. aeolicus that entirely relies on its HARP for RNase P activity, pre-tRNAs apparently coevolved with the enzyme, as 38 of the 44 tRNA transcripts carry an A residue and none a stable G-C or C-G bp immediately upstream of the native cleavage site, two attributes that are clearly favorable for accurate and efficient catalysis by this class of protein-only RNase P.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":"1503-1522"},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439589/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144761163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Two-step target recognition for the competitive inhibition activity of an anti-VEGF aptamer. 抗vegf适配体竞争性抑制活性的两步目标识别。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080524.125

Koh Takeuchi, Takumi Ueda, Misaki Imai, Miwa Fujisaki, Masanari Tsujimura, Yuji Tokunaga, Yutaka Kofuku, Ichio Shimada

{"title":"Two-step target recognition for the competitive inhibition activity of an anti-VEGF aptamer.","authors":"Koh Takeuchi, Takumi Ueda, Misaki Imai, Miwa Fujisaki, Masanari Tsujimura, Yuji Tokunaga, Yutaka Kofuku, Ichio Shimada","doi":"10.1261/rna.080524.125","DOIUrl":"10.1261/rna.080524.125","url":null,"abstract":"The anti-vascular endothelial growth factor (VEGF) aptamer, t44.27, is a 27-mer RNA that functions as the active component of pegaptanib, an antiangiogenic medicine for neovascular age-related macular degeneration. The t44.27 aptamer is extensively 2'-modified and tightly binds to the heparin-binding domain (HDB) of VEGF165 in a Ca2+-dependent manner. However, the molecular mechanism by which the aptamer selectively recognizes VEGF HDB to antagonize its function is poorly understood. We found that t44.27 binds to VEGF HBD in a two-step manner using a different region in the molecule: a transient interaction using a structured region of t44.27 followed by a tight complex formation with a larger interaction surface. Ca2+ binding stabilizes t44.27 base-pair formation suitable for the initial transient interaction. Meanwhile, the tight complex formation was essential for t44.27 to exert a competitive inhibition of heparin binding to VEGF HBD. These results provide structural insight into how the RNA aptamer specifically interacts with its target molecule to inhibit its activity.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":"1368-1378"},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439592/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144761165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

40S ribosomal subunits scan mRNA for the start codon by one-dimensional diffusion. 40S核糖体亚基通过一维扩散扫描mRNA寻找起始密码子。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080493.125

Hironao Wakabayashi, Mingyi Zhu, Elizabeth J Grayhack, David H Mathews, Dmitri N Ermolenko

引用次数: 0

The codon context provides cis-acting immune evasion for the human papilloma virus (HPV) E6. 密码子上下文为人乳头瘤病毒（HPV） E6提供了顺式作用的免疫逃避。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080390.125

Aikaterini Thermou, Chrysoula Daskalogianni, Lixiao Wang, Laurence Malbert-Colas, Van-Trang Dinh, Mathilde Lavigne, Marc Blondel, Robin Fahraeus, Justine Habault

{"title":"The codon context provides cis-acting immune evasion for the human papilloma virus (HPV) E6.","authors":"Aikaterini Thermou, Chrysoula Daskalogianni, Lixiao Wang, Laurence Malbert-Colas, Van-Trang Dinh, Mathilde Lavigne, Marc Blondel, Robin Fahraeus, Justine Habault","doi":"10.1261/rna.080390.125","DOIUrl":"https://doi.org/10.1261/rna.080390.125","url":null,"abstract":"Human papilloma viruses (HPV) are linked to cancers but how virus-carrying tumor cells express HPV-encoded antigens without attracting the immune system is still poorly understood. Here we show how low- and high-risk HPV types equally exploit a cis-acting mechanism to limit the translation of the E6 mRNA, reducing the production of antigenic peptide substrates for the major histocompatibility class I (MHC-I) pathway. Introducing particular combinations of preferable codons throughout the HPV-16 E6 mRNA promotes mRNA translation and production of antigenic peptide substrates in mammalian cells but has minimal impact on E6 synthesis in S. cerevisiae. Using a gradual synonymous codon exchange, we identified a codon series with a significant effect on E6 translation rate. Unexpectedly, changing four non-preferable codons to preferable codons in the wild-type sequence resulted in an approximate 50% reduction in E6 expression. However, five additional changes to preferable codon further upstream shifted this inhibition to a strong induction of E6 expression, while they had no effect when introduced alone. These findings suggest a nuanced relationship between tRNA pools and translation rate, emphasizing how HPV uses codon usage to evade immune detection.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":""},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145076101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

ZYS-1 is not an ADAR1 inhibitor. ZYS-1不是ADAR1抑制剂。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080721.125

Cassandra N Smoak, Estelle N Gardner, Renee N Chua, Kyle A Cottrell

{"title":"ZYS-1 is not an ADAR1 inhibitor.","authors":"Cassandra N Smoak, Estelle N Gardner, Renee N Chua, Kyle A Cottrell","doi":"10.1261/rna.080721.125","DOIUrl":"10.1261/rna.080721.125","url":null,"abstract":"Adenosine deaminase acting on RNA 1 (ADAR1) edits double-stranded RNA (dsRNA) substrates by the deamination of adenosine to inosine in a process known as A-to-I editing. Modulation of ADAR1 expression and editing activity has previously been described to play a role in cancer development and progression, with upregulation of ADAR1 being observed in a range of cancers. Further, depletion of ADAR1 leads to increased sensing of endogenous dsRNAs by dsRNA sensors in cell lines that require ADAR1 for survival, which are termed ADAR1-dependent. The activation of these sensors induces downstream production of type I interferons as well as translational inhibition and apoptosis. Therefore, ADAR1 is a promising oncologic therapeutic target. Recently, the small molecule ZYS-1 has been developed and presented as a direct inhibitor of ADAR1. We performed a series of in vitro and cellular experiments to validate the efficacy and specificity of ZYS-1 as an ADAR1 inhibitor. Evaluating the effect of ZYS-1 on cell viability revealed it to be equally cytotoxic to both ADAR1-dependent and ADAR1-independent cell lines, as well as wildtype and ADAR1 knockout cells. Moreover, ZYS-1 treatment had little effect on activation of PKR or induction of IFN stimulated genes. Importantly, treatment with ZYS-1 did not reduce cellular A-to-I editing for several known ADAR1 editing sites, and did not inhibit in vitro A-to-I editing by recombinant ADAR1. Together, these data indicate that ZYS-1 is not a selective inhibitor of ADAR1.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":""},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145076168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular basis for the interactions of eIF2β with eIF5, eIF2B, and 5MP1 and their regulation by CK2. eIF2β与eIF5、eIF2B和5MP1相互作用的分子基础及CK2对它们的调控。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080652.125

Paul A Wagner, Meimei Song, Paul Doran, Ayşenur Şeker, Ralf Ficner, Bernhard Kuhle, Assen Marintchev

{"title":"Molecular basis for the interactions of eIF2β with eIF5, eIF2B, and 5MP1 and their regulation by CK2.","authors":"Paul A Wagner, Meimei Song, Paul Doran, Ayşenur Şeker, Ralf Ficner, Bernhard Kuhle, Assen Marintchev","doi":"10.1261/rna.080652.125","DOIUrl":"10.1261/rna.080652.125","url":null,"abstract":"The heterotrimeric GTPase eukaryotic translation initiation factor 2 (eIF2) delivers the initiator Met-tRNAi to the ribosomal translation preinitiation complex (PIC). eIF2β has three lysine-rich repeats (K-boxes), important for binding to the GTPase-activating protein eIF5, the guanine nucleotide exchange factor eIF2B, and the regulator eIF5-mimic protein (5MP). Here, we combine X-ray crystallography with NMR to understand the molecular basis and dynamics of these interactions. The crystal structure of yeast eIF5-CTD in complex with eIF2β K-box 3 reveals an extended binding site on eIF2β, far beyond the K-box. We show that eIF2β contains three distinct binding sites, centered on each of the K-boxes, and that human eIF5, eIF2Bε, and 5MP1 can bind to all three sites. Our results reveal how eIF2B speeds up the dissociation of eIF5 from eIF2-GDP to promote nucleotide exchange; and how 5MP1 can destabilize eIF5 binding to eIF2 and the PIC, to promote stringent start codon selection. All these affinities are increased by CK2 phosphomimetic mutations, highlighting the role of CK2 in both remodeling and stabilizing the translation apparatus.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":"1419-1435"},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439599/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144650290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Promiscuous RNA editing in lncRNA roX1 is generally nonadaptive. lncRNA roX1中的混杂RNA编辑通常是非适应性的。

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080591.125

Jiyao Liu, Qiuhua Xie, Wanzhi Cai, Hu Li, Yuange Duan, Ling Ma

{"title":"Promiscuous RNA editing in lncRNA roX1 is generally nonadaptive.","authors":"Jiyao Liu, Qiuhua Xie, Wanzhi Cai, Hu Li, Yuange Duan, Ling Ma","doi":"10.1261/rna.080591.125","DOIUrl":"10.1261/rna.080591.125","url":null,"abstract":"Adaptation of adenosine-to-inosine (A-to-I) RNA editing has only been validated for a few nonsynonymous (recoding) sites, where the editable state confers higher fitness than the uneditable or fully edited state. However, adaptation of noncoding RNA editing is unexplored, limiting our understanding of the significance of post-transcriptional modifications. In this study, we report extensive A-to-I editing in Drosophila lncRNA roX1, a key component of the dosage compensation pathway. Despite dramatically higher roX1 expressions in males compared to females, editing levels show minimum gender specificity, indicating nonselective, promiscuous editing. Cross-sample comparison reveals no correlation between roX1 editing levels and the extent of dosage compensation. Furthermore, Adar mutant male flies do not display abnormal dosage compensation of X chromosomal genes. These findings suggest that rampant RNA editing in roX1 is unlikely to play functional roles in dosage compensation and does not appear to offer a selective advantage over either a genomic G or an uneditable allele. Our results align with the molecular error hypothesis that the adaptive changes represent only a small fraction of the genome. Nevertheless, we do not exclude the possibility that, despite a global nonadaptive signal, individual editing sites in roX1 may still be adaptive, contingent on supporting experimental evidence.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":"1357-1367"},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439590/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144744572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative analysis of convergent and divergent T7 RNA polymerase promoters for the synthesis of dsRNA in vivo and in vitro. 趋同型和发散型T7 RNA聚合酶启动子在体内和体外合成dsRNA的比较分析

IF 5 3区生物学

RNA Pub Date : 2025-09-16 DOI: 10.1261/rna.080556.125

Sebastian J Ross, John A Ray, Peter M Kilby, Mark J Dickman

{"title":"Comparative analysis of convergent and divergent T7 RNA polymerase promoters for the synthesis of dsRNA in vivo and in vitro.","authors":"Sebastian J Ross, John A Ray, Peter M Kilby, Mark J Dickman","doi":"10.1261/rna.080556.125","DOIUrl":"10.1261/rna.080556.125","url":null,"abstract":"Double-stranded RNA plays a key role in various biological processes. The discovery of RNA interference, a gene-silencing mechanism, revolutionized the study of gene function. dsRNA has since been used in novel therapeutics and as an agricultural biocontrol alternative to chemical pesticides. Microbial production typically involves expression systems with convergent T7 promoters. However, convergent transcription from DNA-dependent RNA polymerases can lead to transcriptional interference. In this study, we designed multiple plasmid DNA constructs to investigate the effect of convergent and divergent T7 RNA polymerase production of dsRNA via in vitro transcription and in vivo in Escherichia coli, prior to dsRNA yield quantification and analysis of product quality. We demonstrate that higher yields of larger dsRNA are typically obtained using convergent promoters during in vivo production. A typical fold increase of 2.1 was obtained for dsRNAs >400 bp. However, production of smaller dsRNAs (<250 bp) by divergent promoters resulted in increased yields (2.2-fold). Furthermore, our data demonstrate that in vitro transcription production of dsRNA using divergent T7 promoters results in significantly higher yields of dsRNA, with a maximum fold increase of 6.46. Finally, independent of size, we demonstrate that dsRNA synthesized from DNA templates with multiple transcriptional terminators, compared to run-off transcription, improved the quality and purity of dsRNA due to decreased formation of dsRNA multimers or aggregates. This study demonstrates that optimal production of dsRNAs is not limited to a single method and can be optimized depending on the size of dsRNA, application, yield, and quality required.","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":"1403-1418"},"PeriodicalIF":5.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439596/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144619952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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