MERR APEX-seq protocol for profiling the subcellular nascent transcriptome in mammalian cells.
Biotechnology and Bioengineering
Chemistry
Molecular Biology
Molecular/Chemical Probes
RNAseq
Sequence Analysis
Sequencing
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
17 03 2023
17 03 2023
Historique:
received:
14
07
2022
revised:
01
09
2022
accepted:
04
01
2023
medline:
23
10
2023
pubmed:
1
3
2023
entrez:
28
2
2023
Statut:
ppublish
Résumé
Knowledge about the spatial organization of RNAs in eukaryotic cells is crucial for understanding their functions. Here, we present a detailed MERR APEX-seq protocol to achieve spatiotemporally resolved mapping of the subcellular transcriptome in cultured mammalian cells. This protocol provides detailed description of constructing cell lines stably expressing APEX2, immunofluorescence characterization, MERR APEX labeling, enrichment of biotinylated RNA, library construction and high-throughput sequencing, and MERR APEX-seq data analysis. For complete details on the use and execution of this protocol, please refer to Li et al. (2022).
Identifiants
pubmed: 36853684
pii: S2666-1667(23)00015-1
doi: 10.1016/j.xpro.2023.102057
pmc: PMC9898802
pii:
doi:
Substances chimiques
RNA
63231-63-0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
102057Informations de copyright
Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.