A New Method for Studying RNA-binding Proteins on Specific RNAs

doi:10.21769/BioProtoc.4022

	A New Method for Studying RNA-binding Proteins on Specific RNAs
	Sun, Weiping1,2,3 ; Zhang, Ziheng1,2,3 ; Liu, Ji-Long1 ; Zhuang, Min1
	2021-05-20
发表期刊	BIO-PROTOCOL (IF:1.0[JCR-2023],1.4[5-Year])
EISSN	2331-8325
卷号	11 期号:10
DOI	10.21769/BioProtoc.4022
摘要	Proximity- based protein labeling has been developed to identify protein-nucleic acid interactions. We have reported a novel method termed CRUIS (CRISPR-based RNA-United Interacting System), which captures RNA-protein interactions in living cells by combining the RNA-binding capacity of CRISPR/Cas13 and the proximity-tagging activity of PUP-IT. Enzymatically deactivated Cas13a (dCas13a) is fused to the proximity labeling enzyme PafA. In the presence of a guide RNA, dCas13a binds specific target RNA region, while the fused PafA mediates the labeling of biotin-tagged Pup on proximal proteins. The labeled proteins can be enriched by streptavidin pull-down and identified by mass spectrometry. Here we describe the general procedure for capturing RNA-protein interactions using this method.
关键词	Proximity-based protein labeling CRUIS RNA-binding protein PUP-IT CRISPR
收录类别	ESCI
语种	英语
WOS研究方向	Life Sciences & Biomedicine - Other Topics
WOS类目	Biology
WOS记录号	WOS:000654317100006
出版者	BIO-PROTOCOL
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/126953
专题	生命科学与技术学院_博士生生命科学与技术学院_PI研究组_刘冀珑组生命科学与技术学院_PI研究组_庄敏组
通讯作者	Liu, Ji-Long; Zhuang, Min
作者单位	1.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China; 2.Univ Chinese Acad Sci, Beijing, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Shanghai, Peoples R China
第一作者单位	生命科学与技术学院
通讯作者单位	生命科学与技术学院
第一作者的第一单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Sun, Weiping,Zhang, Ziheng,Liu, Ji-Long,et al. A New Method for Studying RNA-binding Proteins on Specific RNAs[J]. BIO-PROTOCOL,2021,11(10).
APA	Sun, Weiping,Zhang, Ziheng,Liu, Ji-Long,&Zhuang, Min.(2021).A New Method for Studying RNA-binding Proteins on Specific RNAs.BIO-PROTOCOL,11(10).
MLA	Sun, Weiping,et al."A New Method for Studying RNA-binding Proteins on Specific RNAs".BIO-PROTOCOL 11.10(2021).