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

Elena Romano; Piera Trionfini; Roberta Giampietro; Ariela Benigni; Susanna Tomasoni

doi:10.1016/j.scr.2021.102580

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

Stem Cell Res. 2021 Dec:57:102580. doi: 10.1016/j.scr.2021.102580. Epub 2021 Oct 18.

Authors

Elena Romano¹, Piera Trionfini², Roberta Giampietro¹, Ariela Benigni¹, Susanna Tomasoni³

Affiliations

¹ Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
² Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: piera.trionfini@marionegri.it.
³ Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: susanna.tomasoni@marionegri.it.

Abstract

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.