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Merck

Generation of a homozygous CIITA knockout iPS cell line using the CRISPR-Cas9 system.

Stem cell research (2021-10-24)
Elena Romano, Piera Trionfini, Roberta Giampietro, Ariela Benigni, Susanna Tomasoni
RESUMEN

Human induced pluripotent stem cells (iPSCs) have great promise in regenerative medicine. However, several limitations, including immune-incompatibility, have raised concerns regarding their clinical application. Recent studies have shown that human iPSCs and their derivatives lose their immunogenicity when major histocompatibility complex (MHC) class I and II genes are inactivated and CD47 is over-expressed. In this study, we used CRISPR-Cas9 technology to generate an isogenic iPSC line with a homozygous frameshift mutation in the MHC II transactivator (CIITA) gene. The CIITA-/- iPSCs exhibit typical morphology of pluripotent cells, normal karyotype, expression of pluripotency markers and differentiation capacity in the three germ layers.

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Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Anticuerpo anti-actina, αmúsculo liso- Cy3 monoclonal de ratón, clone 1A4, purified from hybridoma cell culture