{"title":"Protocol for forming G-quadruplexes from double-stranded DNA during transcription.","authors":"Jiayu Zhang, Jiayuan Gong, Jun Chen, Hanqing Chen","doi":"10.1016/j.xpro.2025.103677","DOIUrl":null,"url":null,"abstract":"<p><p>G-quadruplexes are four-stranded nucleic acid structures that play vital roles in regulating gene expression, maintaining genomic stability, and supporting various biological processes. This protocol details their formation from double-stranded DNA via in vitro transcription. It includes steps for selecting suitable DNA templates, assembling necessary components (such as RNA polymerase, nucleotides, and buffers), setting optimal incubation conditions, and performing dimethyl sulfate (DMS) footprinting to analyze the structures. For complete details on the use and execution of this protocol, please refer to Zhang et al.<sup>1</sup> and Gong et al.<sup>2</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103677"},"PeriodicalIF":1.3000,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11952794/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2025.103677","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/3/13 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
G-quadruplexes are four-stranded nucleic acid structures that play vital roles in regulating gene expression, maintaining genomic stability, and supporting various biological processes. This protocol details their formation from double-stranded DNA via in vitro transcription. It includes steps for selecting suitable DNA templates, assembling necessary components (such as RNA polymerase, nucleotides, and buffers), setting optimal incubation conditions, and performing dimethyl sulfate (DMS) footprinting to analyze the structures. For complete details on the use and execution of this protocol, please refer to Zhang et al.1 and Gong et al.2.
g -四联体是一种四链核酸结构，在调节基因表达、维持基因组稳定性和支持各种生物过程中发挥重要作用。该方案详细说明了它们的形成从双链DNA通过体外转录。它包括选择合适的DNA模板，组装必要的组件（如RNA聚合酶，核苷酸和缓冲液），设置最佳孵育条件，并执行硫酸二甲酯（DMS）足迹分析结构的步骤。关于本协议使用和执行的完整细节，请参考Zhang et al.1和Gong et al.2。
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