Protocol for forming G-quadruplexes from double-stranded DNA during transcription
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>.
Authors
- Jun Chen (ORCID: https://orcid.org/0000-0001-7637-5982)
- Jiayu Zhang (ORCID: https://orcid.org/0000-0001-6035-479X)
- Hanqing Chen (ORCID: https://orcid.org/0000-0001-5442-2071)
- Jia-yuan Gong
Institutions
- Capital Medical University (CN)
- Chinese Academy of Sciences (CN)
- Institute of Microbiology (CN)
- Institute of High Energy Physics (CN)
- University of Chinese Academy of Sciences (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-03-01
- DOI
- https://doi.org/10.1016/j.xpro.2025.103677
- Primary Topic
- DNA and Nucleic Acid Chemistry
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- Youth Innovation Promotion Association of the Chinese Academy of Sciences
- Natural Science Foundation of Beijing Municipality
- Guangzhou Science and Technology Program key projects
- Institute of High Energy Physics
- Youth Innovation Promotion Association
- Basic and Applied Basic Research Foundation of Guangdong Province