TABLE 2.

Physical delivery of CRISPR/Cas systems.

Approaches	Delivery strategies	CRISPR/Cas systems	Target genes	Target cells/tissues	Species/diseases	References
Microinjection	in vitro	Cas9 mRNA and sgRNAs	Ppar-γ, Rag1	One-cell embryos	Cynomolgus monkey	Niu et al. (2014)
	in vitro	Cas9 mRNA and sgRNAs	APOE, B2M, PRF1, and PRKDC	Fertilized eggs	Rat	Ma et al. (2014)
	in vitro	Cas9 mRNA and sgRNAs	Gal4	Embryos	Zebrafish	Kimura et al. (2014)
	in vitro	Cas9 mRNA and sgRNAs	APOE, CD36, CFTR, LDLR, apoC-III, SCARB1, LEP, LEP-R, RyR2	Embryos	Rabbits	Yang et al. (2014)
	in vitro	Cas9 mRNA and sgRNAs	MSTN	Zygotes	Sheep	Crispo et al. (2015)
	in vitro	Cas9 protein and sgRNA	-	Embryos	Xenopus laevis	Corkins et al. (2022)
Electroporation	in vitro	CRISPR/Cas9 plasmid	APPSwe, PSEN1M146V	iPS cells	Human	Paquet et al. (2016)
	in vitro	CRISPR/Cas9 plasmid	exon 45–55 mutation hotspot region	Myoblasts	DMD	Ousterout et al. (2015)
	ex vivo	CRISPR/Cas9 plasmid	PD-1	Human primary T cells	Cancer	Su et al. (2016)
	ex vivo	Cas9/sgRNA RNP	TGFBR2, HPRT1	Human NK Cells	-	Naeimi et al. (2018)
	In vivo	Cas9/sgRNA RNP	Sox2, GFP	-	Axolotl	Fei et al. (2016)
	in vivo	Cas9/sgRNA RNP	exon80	Skin	Mouse	Wu et al. (2017b)
Hydrodynamic injection	in vivo	CRISPR/Cas9 plasmid	HBV	Hepatoma cells	Transgenic mouse	Zhu et al. (2016)
	in vivo	CRISPR/Cas9 plasmid	Pten, p53	Liver	Mouse	Xue et al. (2014)
Membrane deformation	in vitro	CRISPR/Cas9 plasmid	GFP, NUAK2, AAVS1	293T, MCF7, SUM159, SU-DHL-1, AB2.2	Human, mouse	Han et al. (2015)
Sonoporation	in vitro	Cas9/sgRNA RNP	GFP	B16F10	Mouse	Hansen-Bruhn et al. (2018)
Lance array nanoinjection	in vitro	CRISPR/Cas9 plasmid	GFP	HeLa	Human	Sessions et al. (2016)

Notes: Ppar-γ: peroxisome proliferator-activated receptor; Rag1: recombination activating 1; APOE: apolipoprotein; B2M: beta-2-microglobulin; PRF1: perforin 1; PRKDC: protein kinase, DNA-activated, catalytic subunit; GAL4: galactose-responsive transcription factor; CD36: cluster of differentiation 36; CFTR: cystic fibrosis transmembrane conductance regulator; LDLR: low-density lipoprotein receptor; apoC-III: apolipoprotein C-III; SCARB1: scavenger receptor class B,member1; LEP: leptin; LEP-R: leptin receptor; RyR2: ryanodine receptor 2; MSTN: myostatin; iPS: pluripotent stem cells; APP^Swe: amyloid precursor protein; PSEN1^M146V: presenilin 1; DMD: duchenne muscular dystrophy; PD-1: programmed death-1; TGFBR2: TGF-beta type II, receptor; HPRT1: hypoxanthine phosphoribosyltransferase 1; HBV: hepatitis B virus; GFP: green fluorescent protein; RNP: ribonucleoprotein.