Table 2.

Application of CRISPR as a therapeutic tool for common multifactorial diseases of humans.

Disease	Manipulated gene	Target	Cell type	Species	In vitro/in vivo	Delivery	Outcome	Ref
Cancer	HPV16 genome	Exon 7	HPV positive SiHa and Caski cells, HPV negative C33 A HEK 293	Human	In vitro	Luciferase reporter pSSA Rep3-1 plasmid	Apoptosis and growth inhibition of cells. No inhibition and apoptosis. Downregulation of E7 protein and upregulation of tumor suppression protein pRb.	[65]
	Pten gene and p53 gene simultaneously		Liver cells	Mouse	In vivo	Hydrodynamic injection to deliver of Cas9 and sgRNAs	Liver tumors similar to those caused by Cre-loxP mediated deletion of p53.	[66]
	Pten gene only		Liver cells	Mouse	In vivo		Akt phosphorylation and lipid accumulation phenocopying gene deletion using Cre-loxp deletion.	[66]
	AsLX1 homozygous mutation	Genomic region overlapping AsLX1 mutations observed in KBM5 cells	KBM5 cell line	Mouse	In vivo	pX458 vector transfection	Longer cell survival was observed when compared to cells that were not rectified. Normal cell function rectified and downregulated polycomb repressive complex 2 genes. Longer survival in mice with X enograft of corrected cell lines as opposed to these X enografted with uncorrected cell lines. Correction of driver mutations in leukemia cells increases survival in in vivo mice.	[67]
	CDK 11	4th coding exon of CDK11	KHOS and U 205 osteosarcoma cell lines	Human	In vitro	U6gRNA-cas9-2A-GFP	Decreased viability and proliferation of osteosarcoma cells. Induced apoptosis in KHOS and U20 cell lines. Reduced invasion and migration of cells.	[68]
	MCL 1		Human Burkitt's lymphoma cells. Burkitt's lymphoma xenograft models.	Mouse	In vivo	Dual lentiviral vector system	Apoptosis of lymphoma cells at high frequency. Tumor regression or impaired growth.	[69]
	SGCBP1		MCF-7 and MDA-MB-231 cell lines.	Human	In vitro	Lenti CRISPR/CAS9 vector	Inhibited proliferation of breast cancer cells.	[70]
	KLHDC4	Targeting exon 5 of KLHDC4 gene	Nasopharyngeal carcinoma cells	Mouse	In vivo	pX330 transfection vector	Inhibited growth, migration, cell proliferation, migration of cells, and increased apoptosis.	[71]
Diabetes	INS gene	Exon 2 and exon 3	Porcine primary fibroblast cells	Piglet	In vivo	px458 vector	Successful models generated for study.	[72]
	Letine/leptine receptor genes	Exon 2	C57BL/6J embryo (mice)	Mouse	In vivo	Microinjection of Cas9 mRNA and sgRNAs	Phenotypically identical to mice models involving the use of obese and diabetic mice.	[73]
	Hepatocyte nuclear factor 1B		Human iPSCs	Human	In vitro	Plasmid vectors	It provides extensive insight into the influence that HNF1B knockout mutations can have on the development of diabetes and the molecular mechanisms involved with pancreatic development.	[74]
Cardiovascular diseases	LMNA		1-cell stage zebrafish embryo	Zebrafish	In vivo	Microinjection	Models for the study of early-onset CCD.	[75]
	PCSK9 gene	Exon 1 and exon 2 of the PCSK9 gene	Hepatocytes	Mouse	In vivo	Adenoviral delivery	50% of mice showed loss of function and reduction of LDL levels.	[76]
	ANGPTL3			Mouse	In vivo	Adenoviral vectors	Reduced risk of CHD, reduced blood triglycerides and LDL.	[77]
	ApoE and LDLR gene	Exon 2 of ApoE and LDLR gene		Pig	In vivo	pGL3-U6-gRNA-PGK-puromycin and Cas9 expressing plasmid	Successful generation of pig models.	[78]