. 2020 Apr 27;11(5):473. doi: 10.3390/genes11050473

Table 1.

Studies using the CRISPR technology for functional validation of variants of disease modeling.

Model	Phenotype	Gene	Genomic Target	Aim	Delivery Method	Nuclease	Reference
^H HeLa	adRP	RHO	p.Pro23His	KO	Lipofection (plasmid)	SpCas9	[34]
^H HEK293FT	LCA	CEP290	p.Cys998*	KI	Lipofection (plasmid)	SpCas9	[35]
^H HEK293	USH2/arRP	USH2A	p.Glu767Serfs*21	KI	Lipofection (plasmid)	SpCas9	[36]
^H HEK293	USH2/arRP	USH2A	p.Cys759Phe	KI	Lipofection (plasmid)	SpCas9	[36]
^M 661W	adRP	Rp9	Exon 5	KO	Fugene HD (plasmid)	SpCas9	[37]
^M 661W	adRP	Rp9	p.His137Leu	KI	Fugene HD (plasmid)	SpCas9	[37]
^H hTERT-RPE1	xlRP	RP2	Exon 2	KO	Fugene HD (plasmid)	nCas9 pairs	[38]
	xlRP	RPGR	Exons 2 and 4	KO	Undetermined	uCas9	[39]
	sarRP	PDE6D	Exon 2
	sarRP	INPP5E	Exon 1
	arCORD	RPGRIP1L	Exon 3
^M NSC	arRP	Pde6b	p.Arg560Cys	^C KI	Nucleofection (plasmid)	SpCas9	[40]
rd12 MEFs	LCA	Rpe65	p.Arg44*	^C KI	Electroporation (RNPs)	SpCas9	[41]
PD-Fibroblasts	USH2/arRP	USH2A	p.Glu767Serfs*21	^C KI	Nucleofection (RNPs)	SpCas9	[36]
PD-Keratinocytes	SHRF	EXOSC2	Exon 1 and 4	KO	Lentiviral transduction	SpCas9	[42]
PD-iPSCs	arRP	MAK	c.1513ins353	^C KI	Nucleofection (plasmid)	SpCas9	[43]
	LCA	CEP290	p.Cys998*	KO	Nucleofection (plasmid)	SpCas9
				KO	Electroporation (plasmid)	SaCas9
				^C KI
	adRP	RHO	p.Pro23His	KO
		RHO	p.Pro23His	^C KI	Undetermined (plasmid)	SpCas9
		PRPF31	p.Arg372Glnfs*99	^C KI	Lipofection (plasmid)	SpCas9	[44]
		PRPF31	Exon 7	KO	Nucleofection (plasmid)	SpCas9	[45]
		PRPF8	p.Pro2301Ser	^C KI	Electroporation (gRNA-plasmid and Cas9 mRNA)	Cas9-Gem	[46]
	xlRP	RPGR	p.His562Argfs*20	KI	Electroporation (plasmid)	SpCas9	[47]
	ESCS	NR2E3	p.Val41Alafs*23	KI	Lipofection (plasmid)	SpCas9	[48]
	ESCS	NR2E3	p.Arg73Ser	KI	Electroporation (plasmid)	SpCas9	[48]
	USH2/arRP	USH2A	p.Glu767Serfs*21 p.Cys759Phe	KI	Nucleofection (plasmid)	eSpCas9	[49]
	XLRS	RS1	p.Arg209Cys	KI	Nanodiamonds (linear DNA)	SpCas9	[50]
Caenorhabditis elegans (nematode)	adRP	prp-8	p.Arg2310Gly	KI	Injection (RNPs)	SpCas9	[51]
		prp-8	p.His2309del
		snrp-200	p.Val683Leu
		snrp-200	p.Ser1087Leu
Danio rerio (zebrafish)	arRP	eys	p.Gly1163Valfs*14	KO	Embryo injection (RNPs)	SpCas9	[52]
	arRP	pcare	p.Gly8Glu*19	KO	Embryo injection (RNPs)	SpCas9	[53]
	adRP	rho	p.Cys322Argfs*116	KO	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[54]
	LCA	cct2	p.Leu394His-7del	KO	Embryo injection (RNPs)	SpCas9	[55]
	USH2/arRP	ush2a	p.Cys780Glnfs*32	KO	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[56]
			p.Ala5174*	KO	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[56]
			p.Lys2532Thrfs*56	KI	Embryo injection (RNPs)	SpCas9	[57]
	ESCS	nr2e3	p.Leu162Glnfs*30	KO	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[58]
	arCD	cacna2d4	Undetermined	KO	Embryo injection (RNPs)	SpCas9	[59]
	adFEVR	znf408	p.His455Tyr	KI	Embryo injection (RNPs)	uCas9	[60]
Mus musculus (mouse)	Undetermined	Blimp1	B108 cis-regulatory module	KO	Electroporation—subretinal injection (plasmid)	SpCas9	[61]
	LCA	Cep290	p.Cys998*	KO	AAV transduction (subretinal injection)	SpCas9	[35]
		Cep290	Exon 3	KO	AAV transduction (subretinal injection)	SpCas9	[62]
		Rpe65	p.Asp477Gly	KI	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[63]
		Rpe65	p.Arg44*	^C KI	AAV transduction (subretinal injection)	SpCas9	[41]
		Kcnj13	Exon 2	KO	Zygote injection (Cas9 mRNA and gRNAs)	SpCas9	[64]
	OCA1	Tyr	5’ region	KO	Zygote injection (Cas9 mRNA and gRNAs)	SpCas9	[65]
	Thy1-YFP	YFP	5’ region	KO	AAV transduction (intravitreal injection)	SpCas9	[66]
	arRP	Pde6b	p.Arg560Cys	KI	Electroporation—subretinal injection (plasmid)	SpCas9	[40]
		Pde6b	p.Tyr347Ter	^C KI	Embryo injection (gRNA-plasmid and Cas9 protein)	SpCas9	[67]
		Reep6	p.Leu135Pro	KI	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[68]
		Reep6	Exon 4	KO	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[69]
		Arl2bp	Exon 2	KO	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[70]
		Hkdc1	Exon 2	KO	Undetermined (plasmid)	uCas9	[71]
	adRP	RHO	p.Pro23His	KO	Electroporation (plasmid)	SpCas9	[34]
		RHO	p.Pro23His	KO	Electroporation (plasmid) and AAV transduction (intravitreal injection)	SaCas9 and SaCas9-KKH	[72]
		Rho/RHO	Exon 1	KO	AAV transduction (subretinal injection)	SpCas9	[73]
	arRP/sarRP	Cwc27	p.Lys338Glyfs*25	KO	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[74]
	arRP/adRP	Nrl	Undetermined	KO	AAV transduction (subretinal injection)	SpCas9	[75]
	arRP/adRP	Nrl	Undetermined	KO	AAV transduction (subretinal injection)	SpCas9	[76]
	RD	Slc9a8	Promoter	KO	AAV transduction (subretinal injection)	nmCas9	[77]
	RD	Usp45	Exon 14	KO	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[78]
	adCORD	Gucy2e	Exon 2 and 4	KO	AAV transduction (subretinal injection)	SaCas9	[79]
	Thy1-YFP	YFP	Undetermined	KO	AAV transduction (subretinal injection	SpCas9	[80]
	XLRS	Rs1	p.Arg209Cys	KI	Nanodiamonds—intravitreal injection (linear DNA)	SpCas9	[50]
Rattus norvegicus (rat)	OCA1	Tyr	Exon 2	KO	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[81]
	OCA1	Tyr	p.Arg299His	KI	Embryo injection (Cas9 mRNA and gRNAs)	uCas9	[81]
	adRP	Rho	p.Ser334Ter	KO	Electroporation—subretinal injection (plasmid)	SpCas9	[82]
Xenopus laevis (frog)	adRP	rho	Exon1	KO	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[83]
Xenopus laevis (frog)	adRP	rho	Exon 5	KI	Embryo injection (Cas9 mRNA and gRNAs)	SpCas9	[83]
Drosophila melanogaster (fly)	SHRF	rrp4	Exon 1 and 4	KO	Embryo injection (gRNA-plasmid into Cas9-expressing strain)	SpCas9	[42]
Sus scrofa domesticus (pig)	adRP	RHO	p.Pro23His	KO	AAV transduction (subretinal injection)	SaCas9	[43]
Macaca fascicularis (macaque)	adCORD	GUCY2D	Exon 2 and 4	KO	AAV transduction (subretinal injection)	saCas9	[79]

KO: Knock-Out; KI: Knock-In; MEFs: Mouse Embryonic Fibroblasts; HeLa: Henrietta Lacks Cell Line (Uterine Cell Variety); HEK293: Human Embryonic Kidney 293 Cells; HEK293FT: Fast Growing HEK293 Line Variant; 66W: Immortalized Cone Photoreceptor Cell Line; hTERT-RPE1: Immortalized Retinal Pigment Epithelial Cell Line; NSC: Primary Cultures of Neural Stem Cells; eSpCas9: Enhanced Specificity SpCas9; nCas9: Cas9 Nickase; SpCas9: Cas9 (Streptococcus pyogenes); uCas9: Undetermined Cas9; SaCas9: Cas9 (Staphylococcus aureus); SpCas9-Gem: SpCas9 Fused to Human Geminin Protein; SaCas9-KKH: SaCas9 Recognizing NNNRRT PAMs; nmCas9: Cas9 (Neisseria meningitidis); AAV: Adeno-Associated Virus; PD: Patient-Derived; ad: Autosomal Dominant; ar: Autosomal Recessive; sar: Syndromic Autosomal Recessive; xl: X-Linked; RP: Retinitis Pigmentosa; RD: Retinal Degeneration; CD: Cone Dystrophy; USH2: Usher Syndrome Type 2; XLRS: X-Linked Juvenile Retinoschisis; SHRF: Short Stature, Hearing Loss, Retinitis Pigmentosa and Distinctive Facies Syndrome; FEVR: Familial Exudative Vitreoretinopathy; ESCS: Enhanced S-Cone Syndrome; CORD: Cone-Rod Dystrophy; OCA1: Oculocutaneous Albinism Type 1; LCA: Leber Congenital Amaurosis; Thy1-YFP: Transgenic Mice Expressing Yellow Fluorescent Protein; Symbols: ^H Human-origin cells; ^M Mouse-origin cells; ^C Correction-purpose.