Analyzing efficiency of a lentiviral shRNA knockdown system in human enteroids using western blot and flow cytometry.
STAR Protoc
; 5(2): 103082, 2024 Jun 21.
Article
em En
| MEDLINE
| ID: mdl-38781076
ABSTRACT
Enteroids are in vitro models to study gastrointestinal pathologies and test personalized therapeutics; however, the inherent complexity of enteroids often renders standard gene editing approaches ineffective. Here, we introduce a refined lentiviral transfection protocol, ensuring sufficient lentiviral engagement with enteroids while considering spatiotemporal growth variability throughout the extracellular matrix. Additionally, we highlight a selection process for transduced cells, introduce a protocol to accurately measure transduction efficiency, and explore methodologies to gauge effects of gene knockdown on biological processes.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Western Blotting
/
Lentivirus
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RNA Interferente Pequeno
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Técnicas de Silenciamento de Genes
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Citometria de Fluxo
Limite:
Humans
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2024
Tipo de documento:
Article