Table 1.

Nanoparticle delivery systems for CRISPR-Cas9.

Delivery System	Crispr-Cas Form	Study Objective	Target Gene	In Vitro/ Vivo	Brief Result	Ref
Lipid-based NPs.	Cas9 mRNA and sgRNA	Hepatocytes, C57BL/6 mice	PCSK9	In vivo	The lipid NPs delivered-CRISPR/Cas9 effectively knocked the protein level of PCSK9 in mouse serum down to 20%.	[99]
Lipid-based NPs	Cas9 mRNA and sgRNA	C57BL/6 mice	PTEN PCSK9	In vivo	SORT LNPs mediated effective tissue-specific genes PTEN and PCSK9 editing in the liver.	[100]
Lipid-based NPs	Cas9 mRNA and sgRNA	Hela cells	GFP HPV18E6	In vitro	treatment of HeLa cells with PBA-BADP/Cas9 mRNA/sgHPV18E6 NPs showed GFP knocked out efficiency up to 50% and resulted in 18.7% indel of HPV18E6 gene.	[101]
Lipid-based NPs	Ribonucleoproteins	U87 cells Mice bearing tumor	PLK1	Both in vitro and in vivo	LHNPs co-encapsulated with Cas9 and minicircle sgRNA were capable of efficiently inhibiting PLK1 expression to 36.3% and inhibit tumor growth	[102]
Lipid-based NPs	Cas9 mRNA and sgRNA	BMDMs C57BL/6 mice	NLRP3	Both in vitro and in vivo	Disrupt NLRP3 of macrophages in vitro by CLANmCas9 with an efficiency rate of 70.2%, compared to the rate of 58.6% in vivo.	[106]
Lipid-based NPs	Cas9 mRNA and sgRNA	Splenic endothelial cells HEK293	ICAM-2	In vivo	LNPs can edit endothelial cells successfully, and the ideal Cas9: sgRNA ratio will depend on the relative stability of the two molecules.	[107]
Lipid-based NPs	Cas9 mRNA and sgRNA	HEK293, GBM 005 cells	GFP, PLK1	In vivo	CRISPR-LNPs against PLK1 enabled up to ~70% gene editing in vivo, inhibited tumor growth by 50%, and improved survival by 30%.	[108]
Lipid-based NPs	Ribonucleoproteins	B16F10 cells	PD-L1	In vitro	VLN-sgPD-L1 reduced the expression of PD-L1 to 41.3% and thus suppressed tumor growth in vivo.	[109]
LNP-INT01	Cas9 mRNA and sgRNA	Cd-a-mice	TTR	In vivo	CRISPR-LNPs against TTR in the liver of mice resulted in a 97% reduction in serum protein levels that persisted for at least 12 months.	[110]
Lipidoid NPs	Ribonucleoproteins	Hela-DsRed cells	GFP	In vivo	LNPs-based CRISPR/Cas9 system displayed high GFP knockout efficacies ~70% with low cytotoxicities.	[58]
Gold/lipid NPs	Plasmid DNA	Melanoma	Plk-1	Both in vitro and in vivo	AuNPs-based CRISPR/Cas9 system led to about 65% down-regulation of Plk-1 protein triggered by the photothermal effect.	[111]
Gold NPs	Ribonucleoproteins	Fragile X syndrome	mGluR5	In vivo	CRISPR-Gold targeting the mGluR5 gene reduced the protein level by 40–50% in the mouse models that have fragile X syndrome	[112]
Au NPs	Ribonucleoproteins	HSPCs	CCR5	In vitro	NPs-mediated CRISPR/Cas9 successfully penetrated into HSPCs and produced up to 17.6% total editing.	[113]
Au NPs	Ribonucleoproteins	Hepa 1-6 cells mice	Pcsk9	Both in vitro and in vivo	This Au Nps-based CRISPR/Cas9 delivery system induced significant Pcsk9 editing in vitro and reduced the LDL-C level to 30% compared with the control group by knocking out the Pcsk9 gene in mice.	[114]
polymeric NPs	Plasmid DNA	Chronic myeloid leukemia	CML-related BCR-ABL	Both in vitro and in vivo	CLANpCas9/gBCR-ABL disrupted the BCR-ABL gene in vitro with an efficiency rate of 46.8%, reduced the mRNA level to 41.9%, and greatly inhibited the protein expression of BCR-ABL in CML mice.	[115]
polymeric NPs	Plasmid DNA	HEK293T cell	dTomato	in vitro	Polymeric microcarriers for CRISPR/Cas9 displayed high gene knockout efficiency up to 70% in the transfected cells.	[116]
polymeric NPs	Plasmid DNA	HFD-induced T2D mice	NE	In vivo	CLANpCas9/gNE targeting the neutrophil elastase (NE) gene effectively disrupted the NE gene in the mouse have type 2 diabetes (T2D) with the gene knock-out rate of 26.4% and mitigated the insulin resistance by reducing neutrophils-related inflammation	[117]
polymeric NPs	Plasmid DNA	Hela cells HEK293T cell	GFP iRFP	In vitro	A novel reporter system involving PBAEs-CRISPR carrier for easy detection of gene knockout at one and two genomic sites	[118]
Polymeric NPs	Ribonucleoproteins	S. aureus	MecA	In vitro	Cr-Nanocomplex treatment resulted in a significant inhibition in MRSA growth in the presence of methicillin by disrupting the mecA gene.	[119]
Core-shell NPs with iron oxide core and PEI coating	Plasmid DNA	Porcine fetal fibroblasts	H11	In vitro	Magnetic NPs carrying CRISPR/Cas9 displayed 3.5 times higher efficiency compared to the classic lipofection method	[120]
PEI magnetic NPs	Plasmid DNA	HEK293 cell	TLR-3	In vitro	Magnetic NPs-CRISPR/Cas delivery system enabled site-specific incision with the combination of an inhomogeneous magnetic field.	[121]
PLGA NPs	Ribonucleoproteins	HSPCs	γ-globin gene	In vitro	CRISPR/Cas9-PLGA-NPs-mediated gene inaction ofγ-globin gene in HSPCs led to the increase in the HbF expression (51.7%) in a concentration-dependent manner.	[122]
pH-responsive polymeric NPs	Plasmid DNA	B16F10 cells	Cdk5	Both in vitro and in vivo	The CRISPR/Cas9 encapsulated in nanoparticles specifically knock out the Cyclin-dependent kinase 5 (Cdk5) gene to significantly attenuate the expression of PD-L1 on tumor cells.	[123]