Protocol to study the direct binding of proteins to RNA:DNA hybrids or RNA-DNA chimeras in living cells using cross-linking immunoprecipitation.
STAR Protoc
; 5(3): 103292, 2024 Sep 20.
Article
in En
| MEDLINE
| ID: mdl-39264804
ABSTRACT
RNA-binding proteins (RBPs) are involved in many biological processes. The direct interaction between protein and RNA can be studied using cross-linking immunoprecipitation (CLIP) techniques in living cells. Here, we present a protocol to characterize the direct binding of proteins to RNADNA hybrids or RNA-DNA chimeras in living cells using CLIP. We describe steps for RNA-protein UV-C cross-linking in living cells, isolating RNA-protein complexes, RNA labeling, and extracting nucleic acid. We then detail procedures for nuclease treatment and nucleic acid migration.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA
/
RNA
/
RNA-Binding Proteins
/
Immunoprecipitation
Limits:
Humans
Language:
En
Journal:
STAR Protoc
Year:
2024
Document type:
Article
Affiliation country:
Francia
Country of publication:
Estados Unidos