Generation of an OCT4 reporter human induced pluripotent stem cell line using CRISPR/SpCas9.
Stem Cell Res
; 23: 105-108, 2017 08.
Article
em En
| MEDLINE
| ID: mdl-28925359
ABSTRACT
OCT4 is a crucial transcription factor in the pluripotent stem cell gene regulatory network and an essential factor for pluripotent reprogramming. We engineered the previously reported HEL24.3 hiPSC to generate an OCT4 reporter cell line by knocking-in a T2A nuclear EmGFP reporter cassette before the OCT4 gene STOP codon sequence. To enhance targeted insertion, homologous recombination was stimulated using targeted cutting at the OCT4 STOP codon with CRISPR/SpCas9. This HEL24.3-OCT4-nEmGFP cell line faithfully reports endogenous OCT4 expression, serving as a useful tool to examine temporal changes in OCT4 expression in live cells during hiPSC culture, differentiation and somatic cell reprogramming.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Genes Reporter
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Técnicas de Cultura de Células
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Fator 3 de Transcrição de Octâmero
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Células-Tronco Pluripotentes Induzidas
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Sistemas CRISPR-Cas
Limite:
Humans
Idioma:
En
Revista:
Stem Cell Res
Ano de publicação:
2017
Tipo de documento:
Article