PMID- 31805151
OWN - NLM
STAT- MEDLINE
DCOM- 20200324
LR  - 20200324
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 14
IP  - 12
DP  - 2019
TI  - Generation of targeted homozygosity in the genome of human induced pluripotent 
      stem cells.
PG  - e0225740
LID - 10.1371/journal.pone.0225740 [doi]
LID - e0225740
AB  - 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.
FAU - Yoshimura, Yasuhide
AU  - Yoshimura Y
AD  - Department of Genome Biology, Graduate School of Medicine, Osaka University, 
      Suita, Osaka, Japan.
FAU - Yamanishi, Ayako
AU  - Yamanishi A
AD  - Department of Genome Biology, Graduate School of Medicine, Osaka University, 
      Suita, Osaka, Japan.
FAU - Kamitani, Tomo
AU  - Kamitani T
AD  - Department of Genome Biology, Graduate School of Medicine, Osaka University, 
      Suita, Osaka, Japan.
FAU - Kim, Jin-Soo
AU  - Kim JS
AD  - Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea.
FAU - Takeda, Junji
AU  - Takeda J
AUID- ORCID: 0000-0001-6321-0898
AD  - Department of Genome Biology, Graduate School of Medicine, Osaka University, 
      Suita, Osaka, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191205
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (RNA, Messenger)
RN  - EC 3.6.1.- (Bloom syndrome protein)
RN  - EC 3.6.4.12 (RecQ Helicases)
SB  - IM
MH  - Base Sequence
MH  - Chromosomes, Human, Pair 19/genetics
MH  - Clone Cells
MH  - Crossing Over, Genetic
MH  - DNA Breaks, Double-Stranded
MH  - Gene Expression Regulation
MH  - Genes, Reporter
MH  - *Genome, Human
MH  - Histocompatibility Antigens Class I/genetics
MH  - Homozygote
MH  - Humans
MH  - Induced Pluripotent Stem Cells/*metabolism
MH  - Polymorphism, Single Nucleotide/genetics
MH  - RNA, Messenger/genetics/metabolism
MH  - RecQ Helicases/genetics/metabolism
MH  - Telomere/genetics
PMC - PMC6894808
COIS- The authors have declared that no competing interests exist.
EDAT- 2019/12/06 06:00
MHDA- 2020/03/25 06:00
PMCR- 2019/12/05
CRDT- 2019/12/06 06:00
PHST- 2019/07/11 00:00 [received]
PHST- 2019/11/11 00:00 [accepted]
PHST- 2019/12/06 06:00 [entrez]
PHST- 2019/12/06 06:00 [pubmed]
PHST- 2020/03/25 06:00 [medline]
PHST- 2019/12/05 00:00 [pmc-release]
AID - PONE-D-19-19663 [pii]
AID - 10.1371/journal.pone.0225740 [doi]
PST - epublish
SO  - PLoS One. 2019 Dec 5;14(12):e0225740. doi: 10.1371/journal.pone.0225740. 
      eCollection 2019.