CRISPR-Cas9-Mediated Gene Editing in Human Induced Pluripotent Stem Cells

Abstract

AbstractHuman induced pluripotent stem cell (hiPSCs) and gene editing technologies have become broadly accessible in the last few years and are no longer confined to specialized laboratories. As a result of these developments, both techniques are becoming increasingly prominent in many fields of biomedical research. The use of the CRISPR-Cas9 platform has proven much less labor-intensive compared to alternative platforms for gene editing such as TALENs or ZFNs. However, application of CRISPR-Cas9 in hiPSCs can be cumbersome due to the relatively low efficiency of gene editing in these cells, combined with the requirement of advanced techniques for culturing human iPSCs. Here, we provide protocols for CRISPR-Cas9-mediated gene editing in hiPSCs for the generation of gene knockouts, large deletions, and the introduction of a donor template in a safe harbor.