Mitteaux Jérémie, Monchaud David
Institut de Chimie Moléculaire de l'Université de Bourgogne (ICMUB), CNRS UMR 6302, 9 Avenue Alain Savary, 21078 Dijon, France.
STAR Protoc. 2024 Dec 20;5(4):103480. doi: 10.1016/j.xpro.2024.103480. Epub 2024 Dec 10.
The isolation of G-quadruplexes (G4s) from human cells using specific molecular tools constitutes an invaluable step forward in uncovering the biology of these higher-order DNA and RNA structures. Here, we present an improved version of the G4-RNA precipitation (G4RP) protocol developed to identify RNA G4s from human cancer cells. We describe steps for cell treatment and lysis, chemoprecipitation of G4s using TASQ tools, go/no-go steps, and quantitative reverse-transcription PCR (RT-qPCR) quantification and analysis. For complete details on the use and execution of this protocol, please refer to Mitteaux et al..
利用特定分子工具从人类细胞中分离出G-四链体（G4s），这在揭示这些高阶DNA和RNA结构的生物学特性方面迈出了宝贵的一步。在此，我们展示了一种经过改进的G4-RNA沉淀（G4RP）方案，该方案旨在从人类癌细胞中鉴定RNA G4s。我们描述了细胞处理和裂解步骤、使用TASQ工具对G4s进行化学沉淀、关键步骤以及定量逆转录PCR（RT-qPCR）定量和分析。有关该方案使用和执行的完整详细信息，请参考米托等人的研究。
