RRC ID 58135
Author Morishige S, Mizuno S, Ozawa H, Nakamura T, Mazahery A, Nomura K, Seki R, Mouri F, Osaki K, Yamamura K, Okamura T, Nagafuji K.
Title CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs.
Journal Int J Hematol
Abstract The clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system is an efficient genome-editing tool that holds potential for gene therapy. Here, we report an application of this system for gene repair in hemophilia B (HB) using induced pluripotent stem cells (iPSCs). We prepared targeting plasmids with homology arms containing corrected sequences to repair an in-frame deletion in exon 2 of the factor IX (F9) gene and transfected patient-derived iPSCs with the Cas9 nuclease and a guide RNA expression vector. To validate the expression of corrected F9, we attempted to induce the differentiation of iPSCs toward hepatocyte-like cells (HLCs) in vitro. We successfully repaired a disease-causing mutation in HB in patient-derived iPSCs. The transcription product of corrected F9 was confirmed in HLCs differentiated from gene-corrected iPSCs. Although further research should be undertaken to obtain completely functional hepatocytes with secretion of coagulation factor IX, our study provides a proof-of-principle for HB gene therapy using the CRISPR/Cas9 system.
Volume 111(2)
Pages 225-233
Published 2020-2-1
DOI 10.1007/s12185-019-02765-0
PII 10.1007/s12185-019-02765-0
PMID 31664646
MeSH CRISPR-Cas Systems / genetics* Gene Editing* Genetic Therapy / methods* Hemophilia B / genetics* Hemophilia B / therapy* Humans Induced Pluripotent Stem Cells*
IF 2.245
Times Cited 1
Human and Animal Cells 201B7(HPS0063)