G 型四联体和 R 型环的免疫荧光显微镜。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-01-23 DOI:10.1016/bs.mie.2024.01.008

Giulia Miglietta, Jessica Marinello, Giovanni Capranico

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引用次数: 0

摘要

DNA 和 RNA 之间可以形成多种非 B 二级结构。在本章中，我们将重点讨论 G-四重链（G4）和 R-环，它们在结构上可以产生密切的相互作用。近年来，越来越多的证据表明，它们在体内可以相互产生强烈影响，在正常细胞和癌细胞中都具有生理和病理作用。在这里，我们详细介绍了纯化 BG4 和 S9.6 抗体的特异性和准确性方法，以及它们随后在免疫荧光显微镜中的应用，从而能够对 G4s 和 R 环的程度和定位进行单细胞分析。

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Immunofluorescence microscopy of G-quadruplexes and R-loops.

A large variety of non-B secondary structures can be formed between DNA and RNA. In this chapter, we focus on G-quadruplexes (G4) and R-loops, which can have a close structural interplay. In recent years, increasing evidence pointed to the fact that they can strongly influence each other in vivo, both having physiological and pathological roles in normal and cancer cells. Here, we detail specific and accurate methods for purification of BG4 and S9.6 antibodies, and their subsequent use in immunofluorescence microscopy, enabling single-cell analysis of extent and localization of G4s and R-loops.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.