Table 1 Examples of use of non-viral and viral systems for in vivo delivery of CRISPR based therapeutics

Polymeric nanoparticles

Polyethylene glycol (PEG)- poly(lactic-co-glycolic acid) (PLGA) based cationic lipid-assisted polymeric nanoparticles (CLANs)

All-in-one Cas9-guide RNA plasmid

BCR-ABL gene knockout

Chronic myeloid leukemia (CML) mouse model

[84]

Acid-responsive polycation-fluorinated (ARP-F) nanoparticles

All-in-one Cas9-guide RNA plasmid

BIRC5 gene knockout

A549 (lung cancer) xenografts in nude mice

[85]

Core–shell nanoparticles; core – cationic polyplex of plasmid and phenylboronic acid (PBA)-modified low molecular weight (LMW) polyethylenimine (PEI), shell – 2,3-dimethylmaleic anhydride (DMMA)-modified PEG-b-polylysine

All-in-one plasmid expressing dCas9-VP64 and guide RNA

CRISPRa of miR-524

MDA-MB-231 (breast cancer) xenografts

[86]

Hyaluronic acid (HA) decorated CP/Ad-SS-GD

[Disulfide-bridged biguanidyl

adamantine (Ad-SS-GD) complexed with β-cyclodextrin-conjugated low-molecular-weight polyethyleneimime (CP)]

Cas9 protein, guide RNA (Cas9 RNP)

KRAS (mutant) gene knockout

SW480 (colorectal adenocarcinoma) xenografts in nude mice

[87]

Liposomes and lipid nanoparticles

Exosomes

Separate Cas9 and guide RNA plasmids

PARP1 gene knockout

SKOV3 (ovarian cancer) xenografts in nude mice

[88]

Internalizing RGD and cell penetration peptide (mHph3) coated PEI hydrogel-core 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) liposomes

Cas9 protein and sgPLK1 plasmid

PLK1 gene knockout

U87 (glioma) xenografts

[89]

Folate receptor-targeting DOTAP liposomes

All-in-one Cas9-guide RNA plasmid

DNMT1 gene knockout

SKOV-3 (ovarian cancer) xenografts in nude mice

[90]

Micelles of quaternary

ammonium-terminated poly(propylene oxide) (PPO-NMe3) and amphiphilic Pluronic F127

All-in-one Cas9-guide RNA plasmid

E7 gene knockout

HeLa xenografts in nude mice

[91]

R8-dGR (dual targeting cell penetrating peptide) modified liposomes

Separate Cas9 and guide RNA plasmids

HIF1A gene knockout

BxPC3 (pancreatic cancer) xenografts in nude mice

[92]

PEGylated liposomes

All-in-one Cas9-guide RNA plasmid

HPV16-E7/HPV18-E7 gene knockout

CasKi/HeLa (human papilloma virus 16- and 18-positive respectively) cervical cancer xenografts in Rag1 mice

[93]

Near-infrared (NIR) irradiation sensitive micelles

Cas9 protein, guide RNA (Cas9 RNP)

NRF2 gene knockout

CNE-2 (nasopharyngeal carcinoma) xenografts in nude mice

[94]

Microvesicles

Cas9 protein, guide RNA (Cas9 RNP)

IQGAP1 gene knockout

HepG2 (liver cancer) xenografts in nude mice

[95]

Extracellular vesicles

Cas9 protein, guide RNA (Cas9 RNP)

WNT10B gene knockout

HepG2 (liver cancer) xenografts in nude mice

[96]

Cyclic lipid nanoparticles (different formulations)

5-methoxyuridine modified Cas9 mRNA, modified sgRNA

PLK1 gene knockout

OV8 (ovarian cancer) xenografts in nude mice

[97]

Gold nanoparticles

Core–shell nanoparticles; core – cationic TAT peptide modified gold nanoparticle with Cas9 protein and guide RNA plasmid, shell – cationic liposomes composed of DOTAP, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol, modified with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG

Cas9 protein and sgPLK1 plasmid

PLK1 gene knockout

A375 (melanoma) xenografts in nude mice

[98]

Porous nanoparticles

Single stranded DNA complementary to guide RNA, coated with PEI

Cas9 protein, guide RNA

Reporter gene knockout

U2OS-EGFP xenografts

[99]

Hollow mesoporous silica nanoparticles

All-in-one Cas9-guide RNA plasmid

EGFR gene knockout

H22 isografts in Kunming mice

[100]

Viral vectors

Adenovirus

EGFR gene knockout

H1975/A549 (non-small cell lung cancer) xenografts in nude mice

[101]

CRISPRa of DKK3 gene

PC3 (prostate cancer) xenografts in nude mice

[102]

Adeno-associated virus

HPV16 E6 and E7 gene knockout

HPV16 + anal cancer PDX in NOD SCID mice

[103]

Lentivirus

HIF1A gene knockout

SMMC-7721 (liver cancer) xenografts in nude mice

[104]