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SiCLAT:同时对染色质环和活细胞中的活性转录进行成像

Xin Wan¹, Jie Kong², Xiaodi Hu²

¹State Key Laboratory for Complex, Severe, and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China. xinwan@ibms.pumc.edu.cn.

|January 3, 2025

概括

我们开发了SiCLAT,这是一种用于同时标记DNA和RNA的新老鼠模型. 这种工具可以实时观察活细胞中的基因调节和cis元素相互作用.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
细胞生物学细胞生物学

背景情况:

了解基因调节需要同时可视化DNA和RNA动态.
目前的方法往往缺乏在活细胞内实时跟踪两种核酸类型的能力.

研究的目的:

开发一种用于同时进行DNA和RNA标记的新体内系统.
研究细胞分化过程中的染色质相互作用和基因转录的动态.

主要方法:

用稳定的dCas9-dCas13d磁带生成一个小鼠模型,用于双核酸标记.
应用SiCLAT系统来追踪肌体分化过程中的染色体循环和基因转录.

主要成果:

SiCLAT使dCas9和dCas13蛋白在各种细胞类型中的稳定表达成为可能.
实现了对色素环相互作用和相关基因转录的准确标记.
实时成像ciss元素相互作用和活初级细胞中的基因转录被证明.

结论:

SiCLAT提供了一个强大的工具,可以直接实时观察基因调节机制.
该系统有助于对活细胞中的动态增强剂-促进剂和增强剂-增强剂相互作用进行机制研究.

关键词:

三维基因组是3D的基因组.这是CRISPR成像技术.增强者和促进者的互动.遗传小鼠模型模型活的初级细胞是活的初级细胞.非重复性DNA和RNA成像技术

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Chromatin Immunoprecipitation- ChIP

Chromatin Immunoprecipitation- ChIP

Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...