Table 1. Various Approaches for hPSC Genome Engineering.

hPSC Genome Engineering
	Knock-Out		Knock-In
Approach	INDEL	Early Stop Codon/ Exon Excision	Transgene Insertion	Point Mutation/SNP
Mechanism	NHEJ	HDR	HDR	HDR
Min# gRNAs	1 -3	2	2	2
Donor Plasmid	No	Yes	Yes	Yes

We do not elaborate the details for directed HDR Knock-Out or Point Mutation/SNP in this unit but donor plasmids with intervening in-frame stop codons or lacking critical exons may be designed as needed and applied with these protocols. For more detail on introducing Point Mutation/SNP, some researchers have found single DSBs with high concentration mutant DNA oligonucleotide donors to be effective. Please review Miyaoka et al., 2014 for more detail on that approach.