Conditional Depletion of STN1 in Mouse Embryonic Fibroblasts.
4-Hydroxytamoxifen (4-OHT) treatment
CST
Conditional knockout
Genome maintenance
Mouse embryonic fibroblast (MEFs)
Oxidative damage
STN1
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
20 Apr 2024
20 Apr 2024
Historique:
received:
18
12
2023
revised:
20
03
2024
accepted:
21
03
2024
medline:
30
4
2024
pubmed:
30
4
2024
entrez:
30
4
2024
Statut:
epublish
Résumé
The CTC1-STN1-TEN1 (CST) complex is a single-strand DNA-binding protein complex that plays an important role in genome maintenance in various model eukaryotes. Dysfunction of CST is the underlying cause of the rare genetic disorder known as Coats plus disease. In addition, down regulation of STN1 promotes colorectal cancer development in mice. While prior studies have utilized RNAi to knock down CST components in mammalian cells, this approach is associated with off-target effects. Attempts to employ CRISPR/Cas9-based knockout of CST components in somatic cell lines have been unsuccessful due to CST's indispensable role in DNA replication and cell proliferation. To address these challenges, we outline a novel approach utilizing a Cre-loxP-based conditional knockout in mouse embryonic fibroblasts (MEFs). This method offers an alternative means to investigate the function and characteristics of the CST complex in mammalian systems, potentially shedding new light on its roles in genome maintenance. Key features • Conditional depletion of mammalian STN1 using mouse embryonic fibroblast (MEFs). • Analysis of oxidative damage sensitivity using STN1-depleted MEFs. • This protocol requires
Identifiants
pubmed: 38686350
doi: 10.21769/BioProtoc.4977
pii: e4977
pmc: PMC11056013
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4977Informations de copyright
©Copyright : © 2024 The Authors; This is an open access article under the CC BY-NC license.
Déclaration de conflit d'intérêts
Competing interestsAuthors declare no financial or non-financial competing interests.
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