Generation of a DKK2 homozygous knockout human embryonic stem cell line using the CRISPR/Cas9 system.

Dickkopf-2 (DKK2) is an antagonist of canonical Wnt signaling, which is involved invarious biological processes of development, such as epidermal appendage formation andeye development. To identify underlying effects of DKK2 during embryonic development, we generated a DKK2 homozygous knockout human embryonic stem cell (hESC) line through the CRISPR/Cas9 genome-editing technology. This cell line, which maintains normal stem cell morphology and stably expresses pluripotent markers, could provide an ideal platform for exploring the role of DKK2 in embryonic development. In addition, Zeocin selection combined with tiny clone picking might be a highly efficient way to generate gene-knockout hESC lines.