Table I.

Applications of CRISPR/Cas9 gene editing

Disease	Model	Mutated Gene(s)	Editing Strategy	Delivery Strategy	Reference
Duchenne Muscular Dystrophy	In vivo*	Dmd exon 44	sgRNA recognition of a PAM sequence in exon 45 and generation of insertions and deletions 7 base pairs downstream of the 5′-AG-3′ splice acceptor site	Intraperitoneal injection of single stranded AAV - packaged SpCas9 nuclease with double-stranded AAV - packaged sgRNA	[12]
Beta-Thalassemia/Sickle Cell Anemia	Ex vivo/In vivo**	Hemoglobin β subunit	Genetic editing of autologous CD34+ hematopoietic stem and progenitor cells at the erythroid-specific enhancer region of BCL11A	Electroporation of CD34+ hematopoietic stem and progenitor cells obtained from healthy donors, with CRISPR/Cas9 targeting the BCL11A erythroid-specific enhancer	[16]
Fanconi Anemia (FA)	In vitro/In vivo*	FANCA, FANCB, FANCC, FANCD2	Editing of human FA hematopoietic cells harboring mutations in genes encoding for FA proteins involved in different steps of the FA pathway	Electroporation of a ribonucleoprotein complex composed of the Cas9 protein and either a chemically modified (sgRNA4 containing, 2′-O-methyl 3′phosphorothioate (MS-sgRNA4)) or an in vitro-transcribed sgRNA4 (IVT-RNP4)	[15]
Cystic Fibrosis	In vitro	CFTR	Correction of the CFTR locus by homologous recombination (a donor plasmid encoding wild-type CFTR sequences) in cultured intestinal stem cells of CF patients	Lipofectamine-mediated transfection in patient organoids with a plasmid expressing Cas9 and sgRNA	[24]
Children’s Interstitial Lung Disease	In vivo*	Sftpc	CRISPR/Cas9–induced excision of the mutant SftpcI73T gene can rescue the lung from toxic accumulation of the disease-associated protein	Intra-amniotic injection of Ad vectors containing SpyCas9 and an sgRNA into the amniotic cavity of gestational fetuses	[14]
Cholesterol Metabolism	In vivo*	PCSK9	Introduction of loss-of-function mutations into the endogenous PCSK9 gene	Adenovirus to express Cas9 and a CRISPR guide RNA targeting Pcsk9in mouse liver, where the gene is specifically expressed	[13]
HIV	In vitro	N/A	Simultaneous genome editing of CXCR4 and CCR5 by CRISPR/Cas9 to block HIV-1 infection in primary CD4+ T cells	Lenti-sgR5-Cas9 vector, containing the gRNA targeting CCR5 region, was inserted by the different CXCR4 targeting sgRNAs	[20]
Cancer	In vitro/In vivo*	PLK-1	Therapeutic genome editing of PLK1 to invoke cell cycle arrest and cell death in dividing cells	Intraperitoneal injection of lipid nanoparticles with an antibody to an overexpressed receptor on ovarian cancer cells (EGFR) to cause their selective uptake into ovarian tumors	[25]
Type 2 Diabetes Mellitus	In vivo*	N/A	Downregulation of dipeptidyl peptidase-4 (DPP-4) to prevent degradation of glucagon-like peptide-1 (GLP-1)	Delivered ribonucleoprotein designed to edit the DPP-4 gene delivered in a lecithin-based liposomal nanocarrier	[26]
Chronic Myeloid Leukemia (CML)	In vitro/In vivo*	Chimeric BCR-ABL	Disruption of the fusion region of the BCR-ABL gene	Delivered cationic lipid-assisted nanoparticles with a plasmid encoding SpCas9 and guide RNA against BCR-ABL fusion gene	[27]

^*Mouse Model

^**Clinical Trial

PAM – Protospacer Adjacent Motif; sgRNA – single guide RNA; AAV – Adeno Associated Virus