Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines.
Cancer
Cell Biology
Cell-based Assays
Microscopy
Molecular/Chemical Probes
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
17 06 2022
17 06 2022
Historique:
entrez:
27
5
2022
pubmed:
28
5
2022
medline:
31
5
2022
Statut:
epublish
Résumé
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).
Identifiants
pubmed: 35620072
doi: 10.1016/j.xpro.2022.101340
pii: S2666-1667(22)00220-9
pmc: PMC9127197
doi:
Substances chimiques
Antibodies
0
RNA, Messenger
0
RNA-Binding Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
101340Subventions
Organisme : NCI NIH HHS
ID : R01 CA229907
Pays : United States
Informations de copyright
© 2022 The Author(s).
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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