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김진수
유전체 교정 연구단
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Generation of targeted homozygosity in the genome of human induced pluripotent stem cells

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Title
Generation of targeted homozygosity in the genome of human induced pluripotent stem cells
Author(s)
Yoshimura Y.; Yamanishi A.; Kamitani T.; Jin-Soo Kim; Takeda J.
Publication Date
2019-12
Journal
PLOS ONE, v.14, no.12, pp.0225740 -
Publisher
PUBLIC LIBRARY SCIENCE
Abstract
© 2019 Yoshimura et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.When loss of heterozygosity (LOH) is correlated with loss or gain of a disease phenotype, it is often necessary to identify which gene or genes are involved. Here, we developed a region-specific LOH-inducing system based on mitotic crossover in human induced pluripotent stem cells (hiPSCs). We first tested our system on chromosome 19. To detect homozygous clones generated by LOH, a positive selection cassette was inserted at the AASV1 locus of chromosome 19. LOHs were generated by the combination of allele-specific double- stranded DNA breaks introduced by CRISPR/Cas9 and suppression of Bloom syndrome (BLM) gene expression by the Tet-Off system. The BLM protein inhibitor ML216 exhibited a similar crossover efficiency and distribution of crossover sites. We next applied this system to the short arm of chromosome 6, where human leukocyte antigen (HLA) loci are located. Genotyping and flow cytometric analysis demonstrated that LOHs associated with chromosomal crossover occurred at the expected positions. Although careful examination of HLA-homozygous hiPSCs generated from parental cells is needed for cancer predisposition and effectiveness of differentiation, they may help to mitigate the current shortcoming of hiPSC-based transplantation related to the immunological differences between the donor and host
URI
https://pr.ibs.re.kr/handle/8788114/6705
ISSN
1932-6203
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
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