ABSTRACT
Gene editing using custom-designed nucleases in human pluripotent stem cells (PSCs) is a powerful tool for disease modeling and repair and will likely have widespread implications for medical research. There are number of different nuclease systems used for genome editing. The most common ones include zinc finger nucleases, transcription activator-like effector nucleases, and RNA-guided engineered nucleases derived from the bacterial clustered regularly interspaced short palindromic repeat (CRISPR)–Cas9. However, these nucleases differ in several ways, including composition, targetable sites, specificities, and mutation signatures. CRISPR–Cas9 enables targeted genetic modifications in cultured cells and in whole animals and plants. This technology has the ability to perform site-specific deoxyribonucleic acid (DNA) cleavage in the genome and repair which allows high-precision genome editing. A number of different methods have been attempted to deliver the guide RNA s and the Cas9 to human PSCs including DNA, RNA, protein, and viral methods.
