Abstract

Modeling human disease using human pluripotent stem cells (hPSCs), in combination with CRISPR-Cas9 gene editing, are emerging as important strategies for studying mechanisms of pathogenesis. We used the ArciTect™ CRISPR-Cas9 system to generate clones harboring a C-terminus truncation of CDK5 regulatory subunit-associated protein 2 (CDK5RAP2), a gene associated with the development of primary microcephaly (Lancaster et al, Nature 2013). We successfully generated stable clones which we further characterized for cell quality attributes (karyotype, pluripotency, morphology, and marker expression) prior to differentiation into cerebral organoids using STEMdiff™ Human Cerebral Organoid Kit.