PMID- 33945142
OWN - NLM
STAT- MEDLINE
DCOM- 20220623
LR  - 20220627
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2454
DP  - 2022
TI  - CRISPR/Cas9 Ribonucleoprotein Complex-Mediated Efficient B2M Knockout in Human 
      Induced Pluripotent Stem Cells (iPSCs).
PG  - 607-624
LID - 10.1007/7651_2021_352 [doi]
AB  - Advances in induced pluripotent stem cell (iPSC) technology provide a renewable 
      source of cells for tissue regeneration and therefore hold great promise for cell 
      replacement therapy. However, immune rejection of allograft due to human 
      leukocyte antigen (HLA) mismatching remains a major challenge. Considerable 
      efforts have been devoted to overcoming the immunogenicity of allograft 
      transplantation. One of the approaches is an elimination of HLA molecules on the 
      surface of allogeneic cells using genome editing technology to generate universal 
      stem cells. Here, we present a simple and effective genome editing approach to 
      knockout the beta-2-immunoglobulin (B2M) gene, which encodes B2M protein that forms 
      a heterodimer with HLA class I proteins, in induced pluripotent stem cells 
      (iPSCs) leading to HLA class I (HLA-I) depletion. We also describe detailed 
      procedures for validation of the B2M-knockout iPSCs using flow cytometry, and 
      genotypic analysis for potential off-target regions. Our protocol is also 
      applicable for knocking out other genes in iPSCs and other cell types.
CI  - (c) 2021. Springer Science+Business Media, LLC.
FAU - Thongsin, Nontaphat
AU  - Thongsin N
AD  - Siriraj Center for Regenerative Medicine, Research Department, Faculty of 
      Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
AD  - Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol 
      University, Bangkok, Thailand.
FAU - Wattanapanitch, Methichit
AU  - Wattanapanitch M
AD  - Siriraj Center for Regenerative Medicine, Research Department, Faculty of 
      Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. 
      methichit.wat@mahidol.ac.th.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (HLA Antigens)
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (Ribonucleoproteins)
SB  - IM
MH  - CRISPR-Cas Systems
MH  - Gene Editing
MH  - HLA Antigens/genetics
MH  - Histocompatibility Antigens Class I/metabolism
MH  - Humans
MH  - *Induced Pluripotent Stem Cells
MH  - Ribonucleoproteins/metabolism
OTO - NOTNLM
OT  - CRISPR/Cas9
OT  - Gene knockout
OT  - HLA-I-null iPSCs
OT  - Human leukocyte antigens (HLAs)
OT  - Induced pluripotent stem cells (iPSCs)
OT  - Universal stem cells
EDAT- 2021/05/05 06:00
MHDA- 2022/06/24 06:00
CRDT- 2021/05/04 12:41
PHST- 2021/05/05 06:00 [pubmed]
PHST- 2022/06/24 06:00 [medline]
PHST- 2021/05/04 12:41 [entrez]
AID - 10.1007/7651_2021_352 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2022;2454:607-624. doi: 10.1007/7651_2021_352.