Generation of Knockout Human PSCs Using CRISPR/Cas9 Editing with a Donor Template

Human pluripotent stem cells (hPSCs), which include human embryonic stem cells and human induced pluripotent stem cells, offer tremendous potential for studying early human development, modeling human diseases, and screening drugs. By incorporating various genome-editing technologies into hPSCs, we can explore the roles of genes crucial for embryonic development, assess the effect of genetic mutations in human diseases, and identify the genes that can be targeted by drugs in a high-throughput manner. Here, we focus on cluster regularly interspaced palindromic repeats (CRISPR)/Cas9-meditated gene knockout technologies in hPSCs, which serve as valuable tools for investigating gene functions in cell fate decisions of hPSCs, including pluripotency maintenance and lineage differentiation.