Gene Editing in Pluripotent Stem Cells and Their Derived Organoids

Table 2

Landmarks and trends of gene editing in life and medical sciences.

	Editing methods	Target cells	Targeted genes	Virus transfection	Animal models	Points	Year
MegN	KI	mESCs	Villin locus	Yes	/	Induction of gene-targeting and homologous recombination events	1998 [45]
	M	293T	RAG1 locus	No	/	Targeting endogenous genes; low targeting efficiency; with cytotoxicity	2009 [46]
	KI	293T	π10 locus	Yes	/	Delivering meganucleases into cells in a transient and dose-controlled manner; low targeting efficiency; with cytotoxicity	2011 [47]
ZFN	KI/GFP	hESCs	OCT4 locus、AAVS1 locus	Yes	m	Gene targeting in hESCs	2009 [48]
	M	hESCs	Genomic α-synuclein locus (SNCA)	No	/	Genome editing in hESCs; off-target detection needs to be improved; targeting efficiency needs to be enhanced	2011 [49]
	KO	hiPSCs	LRRK2 (sigma)	No	m	Parkinson’s pathogenesis; patient-derived iPSCs; low targeting efficiency; with cytotoxicity	2013 [50]
	M	hiPSCs	MAPT	Yes	/	Designed mutation iPSCs; FTD pathogenesis; targeting efficiency needs to be enhanced	2018 [51]
	KO	FRT cells	CFTR	No	r	Disease targets; designed KO model	2020 [52]
TALEN	KI/GFP	hPSCs	OCT4 locus	No	/	Genetic engineering for hPSCs; targeting efficiency like ZFN	2011 [53]
	KO	hiPSCs	TNNT2, LMNA/C, TBX5, MYH7, ANKRD1NKX2.5	Yes	/	Human-based KO cell model in vitro; greater freedom and flexibility in target site selection than CRISPR	2017 [36]
	KI/GFP	hiPSCs	AAVS1 locus	No	/	3D organoid models; GFP; mechanistic studies	2019 [23]
CRISPR	KO	hiPSCs	AAVS1 safe harbor locus	No	/	TetO inducible system; feasibility and reversibility of CRISPRi; high off-target efficiency	2016 [54]
	M	hiPSC	FBN1	No	/	Vascular models, human iPSCs; pathogenesis of MFS	2017 [55]
	KI/GFP	hESCs	gRNAs made from the lentiGuide-puro construct	Yes	/	A genome-scale screening; hESCs; impaired differentiation	2019 [56]
	KO	hPSCs	NRL	No	/	A 3D organoid model; disease pathogenesis; high targeting efficiency	2021 [43]

ESCs: embryonic stem cells; FTD: frontotemporal dementia; FRT: Fischer rat thyroid; h: human; iPSCs: induced pluripotent stem cells; KI: knockin; KO: knockout; m: mouse; M: mutation; MFS: Marfan syndrome; PSCs: ESCs and iPSCs; r: rat.