Journal
NUCLEIC ACIDS RESEARCH
Volume 45, Issue 9, Pages 5198-5207Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkx130
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Funding
- Japan Society for the Promotion of Science (JSPS) KAKENHI [10759509]
- Keihanshin Consortium for Fostering the Next Generation of Global Leaders in Research (K-CONNEX)
- Human Resource Development Program for Science and Technology, MEXT
- Japan Agency for Medical Research and development, AMED [15fk0210002h0002, 15bk0104005h0003]
- Grants-in-Aid for Scientific Research [16K18478, 15H06365] Funding Source: KAKEN
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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.
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