CRISPR/Cas9-engineered human ES cells harboring heterozygous and homozygous c-KIT knockout

含有杂合和纯合 c-KIT 敲除基因的 CRISPR/Cas9 工程化人类 ES 细胞

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作者:Marcelo A S de Toledo, Xuhuang Fu, Frederick Kluge, Katrin Götz, Susanne Schmitz, Paul Wanek, Herdit M Schüler, Kristina Pannen, Nicolas Chatain, Steffen Koschmieder, Tim H Brümmendorf, Martin Zenke

Abstract

The receptor tyrosine kinase c-KIT (CD117) has a key role in hematopoiesis and is a marker for endothelial and cardiac progenitor cells. In vivo, deficiency of c-KIT is lethal and therefore using CRISPR/Cas9 editing we generated heterozygous and homozygous c-KIT knockout human embryonic stem cell (ES cell) lines. The c-KIT knockout left ES cell pluripotency unaffected as shown by immunofluorescence and trilineage differentiation potential. Heterozygous and homozygous c-KIT knockouts showed complete loss of exon 17, resulting in ablation of c-KIT protein from the cell surface. c-KIT knockout ES cells provide a valuable tool for further investigating c-KIT biology.

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