Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines. STAR Protoc 2022 Jun 17;3(2):101340
Date
05/28/2022Pubmed ID
35620072Pubmed Central ID
PMC9127197DOI
10.1016/j.xpro.2022.101340Scopus ID
2-s2.0-85130357879 (requires institutional sign-in at Scopus site) 6 CitationsAbstract
Conventional proximity ligation assay (PLA) suffers from target specificity issues that curtail their accuracy on interpreting proximal interactions in cell biology. Here, we present a reliable and sensitive approach by including a fluorochrome-labeled mRNA fragment along with biotin-labeled RNA probe and a target-specific antibody, which were used to generate proximal ligation signals through linear connectors in intact cells. This protocol will be particularly useful for studying the proximal interactions between RNA binding proteins (RBPs) and their target mRNAs in cells. For complete details on the use and execution of this protocol, please refer to George et al. (2021).
Author List
George J, Mittal S, Kadamberi IP, Pradeep S, Chaluvally-Raghavan PAuthors
Pradeep Chaluvally-Raghavan PhD Associate Professor in the Obstetrics and Gynecology department at Medical College of WisconsinSunila Pradeep PhD Associate Professor in the Obstetrics and Gynecology department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AntibodiesBiophysical Phenomena
Cell Line
RNA, Messenger
RNA-Binding Proteins
