Highly efficient biallelic genome editing of human ES/iPS cells using a CRISPR/Cas9 or TALEN system.
Nucleic Acids Res
; 45(9): 5198-5207, 2017 May 19.
Article
en En
| MEDLINE
| ID: mdl-28334759
ABSTRACT
Genome editing research of human ES/iPS cells has been accelerated by clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) and transcription activator-like effector nucleases (TALEN) technologies. However, the efficiency of biallelic genetic engineering in transcriptionally inactive genes is still low, unlike that in transcriptionally active genes. To enhance the biallelic homologous recombination efficiency in human ES/iPS cells, we performed screenings of accessorial genes and compounds. We found that RAD51 overexpression and valproic acid treatment enhanced biallelic-targeting efficiency in human ES/iPS cells regardless of the transcriptional activity of the targeted locus. Importantly, RAD51 overexpression and valproic acid treatment synergistically increased the biallelic homologous recombination efficiency. Our findings would facilitate genome editing study using human ES/iPS cells.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Genoma Humano
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Alelos
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Células Madre Pluripotentes Inducidas
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Sistemas CRISPR-Cas
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Células Madre Embrionarias Humanas
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Nucleasas de los Efectores Tipo Activadores de la Transcripción
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Edición Génica
Límite:
Humans
Idioma:
En
Revista:
Nucleic Acids Res
Año:
2017
Tipo del documento:
Article
País de afiliación:
Japón