Table 1.

In vivo CRISPR-Cas strategies demonstrating functional rescue in animal models

Organ	Human disease	Editing strategy	Disease model	Administration	Ref.
Eye	Retinitis pigmentosa, autosomal recessive, PDE6B mutation	SpCas9/RecA mediated HDR	Pde6b Y347X, nonsense mutation mice	SpCas9, sgRNA, RecA-MS2 Plasmid and ssDNA donor, subretinal and electroporation	244
	Retinitis pigmentosa, autosomal dominant • P23H mutation (class 2) • Class 1	Knockdown Rho	Rho-P23H transgenic mice	Cas9-sgRNA plasmids, subretinal and electroporation	245
		Knockdown mutant Rho	Rho-P23H heterozygous mice	SpCas9-VRQR plasmid and sgRNA plasmid, subretinal and electroporation	143
			Rho S334ter rat	Cas9-sgRNA plasmid, subretinal and electroporation	141
		Knockdown of transcription factor Neural Leuzine zipper (Nrl)	Rho−/−, Rd10, Rho-P347S mice	AAV8-RK-SpCas9, AAV-sgRNA, subretinal	246
	LCA, autosomal recessive • Type 10, IVS26 mutation in CEP20 • Type 2, RPE65 mutation	NHEJ in-frame indels or HR	Humanized CEP290 IVS26 knock-in mice, NHP	AAV5-sgRNA-GRK1-SaCas9, subretinal	222
			Rpe65 nonsense mutation rd12 mice	AAV9-EFS-SpCas9, AAV9-sgRNA-Rpe65-donor, subretinal	247
	Meesman epithelial corneal dystrophy, dominant negative, KRT12 mutation	Knockdown, allele specific, Deplete mutant KRT12	KRT12-L132P humanized heterozygous mice	SpCas9 and sgRNA plasmids, Intrastromal ocular	142
	Neovascular age-related macular degeneration	Knockdown, NHEJ mediated disruption of Vegfa or Hif1a	Laser-induced choroidal neovascularization in mouse eyes	AAV8-CMV-SpCas9, AAV8-sgRNA, subretinal	146
				AAV9-EFS-LbCpf1-crRNA, intravitreal	248
Neurological	Huntington's disease (CAG trinucelotide), expansion mutant HTT	Knockdown HTT	HD140Q-knock-in mice, human HTT exon 1 with 140 CAG repeats (homozygous and heterozygous)	AAV-MECP2-Cas9, AAV-gRNA, intracranial	249
		Knockdown mutant HTT	R6/2 mice, transgenic mice, human HTT exon 1 with 115–150 CAG repeats	AAV1-hSyn-SaCas9-sgRNA, intracranial	196
	ALS, autosomal dominant, SOD1 mutations	Knockdown human SOD1	Transgenic human SOD1 G93A mutation ALS neonatal mice	AAV9-CMV-SaCas9-sgRNA, intravenous or intracerebroventricular	193,194
		BE3 disruption of SOD1		AAV9-CAG-N-Int-CBE-U6-sgRNA, AAV9-CAG-C-Int-CBE-U6-sgRNA, inthrathecal	195
	Fragile X syndrome	Knockdown GRM5	Fmr1 KO mice, FXS mice model	CRISPR Gold nanoparticle Cas9 RNP and donor DNA, intracranial	82
	Alzheimer's disease	Knockdown Bace1	Five familial Alzheimer's disease transgenic mice, APP knock-in Alzheimer's disease mice	Amphiplilic R7L10 peptide nanocomplex Cas9 RNP, intracranial	7
	Dravet Syndrome, haploinsufficiency, SCN1A loss of function mutation	dCas9-Vp64 mediated Scn1a gene activation (both WT and mutant)	Scn1a heterozygous Dravet Syndrome mice	AAV9-dCas9-VP64, AAV9-sgRNA, intracerebroventricular	250
	Severe obesity, haploinsufficiency, SIM1 or MC4R	dCas9-Vp64 mediated gene activation of Sim1	Sim1/Mc4r heterozygous mice	AAVDJ-CMV-dCas9-Vp64, AAVDJ-sgRNA, hypothalamic	251
Liver	Hemophilia B, X-linked recessive	HDR, insert hFIX-padua exon 2–8 in mFIX exon 2	FIX Knockout mice	AAV8-Cas9, AAV8-sgRNA-Donor DNA	165
		HDR, insert into mFIX into murine ROSA26 safe harbor	R333Q Hemophilia mice	Ad5-Cas9-gRNA, Ad5-EF1α-mFIX, intravenous	252
	Hemophilia B, Y371D mutation in FIX	HDR	Y381D Hemophilia B mice	Naked DNA, SpCas9, ssODN-HDR Donor, intravenous	253
	Hemophilia A, X-linked recessive	Insert human B-domain deleted FVIII in intron 13 of liver-specific albumin locus	FVIII Knockout mice	AAV8-SaCas9-sgRNA, AAV8-donor DNA	166
	OTC deficiency, X-linked recessive	HDR, correct point mutation	spf/ash heterozygous OTC neonate mice	AAV8-TBG-SaCas9, AAV8-sgRNA-donor DNA, intravenous	168
		HDR, insert codon-optimized human OTC into intron 4 of mouse OTC locus		AAV8-TBG-SaCas9, AAV8-sgRNA-TBG-hOTCco, intravenous injection	167
	Hypercholesteolemia	BE3, disruption of mouse W159 Pcsk9	WT C57BL/6 mice	Ad-BE3-sgRNA, retro-orbital	242
		BE3, disruption of human W159 PCSK10 or NHEJ-mediated disruption of PCSK10	Humanized PCSK9 knock-in mice	Ad5-Cas9-sgRNA or Ad5-BE3-sgRNA, Intravenous	243
	Atherogenic dyslipidemia, homozygous familial hypercholesterolemia	BE3, disruption of Q135 Angptl3	Hyperlipidemic Ldlr-knockout mice	Ad-BE3-sgRNA	254
	Phenylketonuria, autosomal recessive	BE3	Homozygous point mutation in Pah-F263S, hyperphenylalaninemia mice	AAV8.N-int-BE3 and AAV8.C-int-BE3, intravenous	255
	Hereditary tyrosinemia type I, autosomal recessive, FAH loss of function mutation	HDR	Fah homozygous mutation in exon 8 mice	SpCas9 mRNA by LNP and AAV8-sgRNA-donor template, intravenous	78
		HDR with nCas9	Homozygous 10-bp deletion in exon 2 of Fah in rat HTI model	Ad-nCas9 and Ad-donor template, intravenous	256
		Base editing, ABE6.3, tRNA nCas9-adenosine deaminase	Fah homozygous mutation in exon 8 mice	Plasmid by hydrodynamic injection or mRNA/sgRNA by LNP, intravenous	257
	Hereditary tyrosinemia type I, autosomal recessive, compound heterozygous mutations in FAH	Allelic exchange by targeting intron 7, NHEJ	compound heterozygous exon 5 insertion/exon 8 mutation in Fah, HTI mouse model	AAV9-SpCas9 and scAAV-sgRNA, intravenous	258
Muscle	DMD, exon 50	Reframing or Exon skipping	ΔEx50 C57/BL6J mice	AAV9-CK8-SpCas9 and AAV9-sgRNA-51, intraperitoneal	259
			ΔEx50 dogs	AAV9-CK8-SpCas9 and AAV9-sgRNA-51, intravenous	125
	DMD, exon 44	Reframing or Exon skipping	ΔEx44 C57BL/6 mice	AAV9-CK8-SpCas9 and AAV9-sgRNA, intraperitoneal	260
				AAV9-CK8-SpCas9 and scAAV-sgRNA	126
	DMD, exons 45–55	Excise exon 52 and 53	mdx4cw mice, nonsense mutation in exon 53	AAV6-CK8-SpCas9 and AAV6-sgRNA or AAV6-CK8-SaCas9-sgRNA, retro-orbital	120
	DMD, nonsense mutation in exon 23	Excise exon 23	Mdx mice, pt mutation in exon 23	AAV9-CMV-SaCas9 and AAV9-sgRNA	119
				AAV8-CMV-SaCas9 and AAV8-sgRNA, Intravenous	124,261
				AAV9-CMV-SaCas9 and AAV9-sgRNA, intravenous	262
				AAV9-miniCMV-spCas9 and AAV9-sgRNA, intraperitoneal	263
		Excise exon 21–23		Ad-Cas9-sgRNA	264
		HDR		CRISPR Gold nanoparticle Cas9 RNP and donor DNA, intramuscular	80
		NHEJ, indels to result in reframing	DMD KO C57/BL6J mice, ΔEx23	AAV9-Spc5-12-CjCas9-sgRNA, intramuscular	265
	Wolff-Parkinson-White syndrome, autosomal dominant inherited disease, H530R mutation in PRKAG2	Disrupt mutant allele	Heart specific H530R transgenic mice, Heterogeneous H530R PRKAG2 knock-in mice	AAV9-Cas9, scAAV9-sgRNA, intravenous or intraventricular	8
Ear	Dominant progressive hearing loss (DFN36), TMC1	Knockdown mutant Tmc1	Beethoven mice, heterozygous missense mutation in Tmc1	Cas9-sgRNA RNP, cationic lipid complex, inner ear	266
				Anc80L65-CMV-SaCas9-sgRNA, inner ear	267
Multiorgan	MPS type I, autosomal recessive	HDR, insert Idua into murine ROSA26 safe harbor	Idua-knock-out MPS I mice	PrecisionX CRISPR/Cas9 SmartNuclease™, Idua donor vector, cationic liposomes, intravenous	182

AAV, adeno-associated virus; ALS, amyotrophic lateral sclerosis; APP, amyloid precursor protein; Bace1, beta-secretase 1; DMD, Duchenne muscular dystrophy; FIX, factor IX; IDUA, α-l-iduronidase; LCA, Leber congenital amaurosis; LNP, lipid nanoparticle; MPS, mucopolysaccharidosis; NHEJ, nonhomologous end-joining; NHP, nonhuman primate; OTC, ornithine transcarbamylase; RNP, ribonucleoprotein; scAAV, self-complementary AAV.