Inactivation of APOBEC3G gene in breast cancer cells using the CRISPR/Cas9 system

CRISPR/Cas9: A powerful genome editing technique for the treatment of cancer cells with present challenges and future directions

“…They reported that both BRCA1m and PARP1m, triple-negative breast cancer (TNBC), cell lines were more sensitive with three different chemotherapeutic drugs, including docetaxel, gemcitabine, and doxorubicin in the 2D tumor culture model as compared to 3D tumor-chip model. Similarly, another study revealed that the non-viral delivery of sgRNA (a component of CRISPR) into HCC1806 and MCF10A cell lines significantly knockout both alleles of APOBEC3G genes in the treatment of breast cancer via blocking the conversion of the G1 to S phase in the cell cycle and inhibiting the cell proliferation [ 43 ]. Yang and his research group exposed that CRISPR/Cas9 technology successfully deleted the individual and co-knockout of CXCR7 and CXCR4 genes (both alleles) in the TNBC cell line (MDA-MB-231) during the treatment of breast cancer by suppressing the tumor cell proliferation, invasion, tumor growth, and invasion [ 44 ].…”

“… Cancer type Target choice Cell line/gene Study type Vector Screening/verification CRISPR effect Mechanism Reference Breast cancer TNBC MDA-MB-231 In vitro Lentiviral Western blotting, PCR Knockout of the CXCR4 or CXCR7 gene Delay the conversion of the G1/S cycle, inhibit cell proliferation, invasion, and mitigation [ 44 ] BRCA1 wild-type, BRCA1m MDA-MB-231, MDA-MB-436 In vitro Plasmid (Sanger sequencing) PCR and T7EI assay Knockout PARP1 inhibitors Apoptosis [ 42 ] PP2A-B55 Cal-51 In vivo Lentiviral Flow cytometry Knock-out of both alleles Inhibit cell proliferation and tumor suppression [ 47 ] Nutlin-3a (organoid) 16 PM0462, 16 PM0408, 8 PM0050 In vivo Lentiviral Western blot analysis Knock-out of PTEN , NF1 , P53 , and RB1 Induction of cellular senescence and cell proliferation [ 108 ] miR-23b, miR27b MCF7 cells In vitro Lentiviral DNA sequencing, qRT-PCR, MTS growth assay Knockout of miR-23b and miR-27b Inhibit cell proliferation and promote migration in miR-27b depleted cells [ 45 ] APOBEC3G MCF10A, HCC1806 In vitro Plasmid Sanger sequencing. Knock-out of both alleles of APOBEC3 Inhibit cell proliferation [ 43 ] Sox2, Sox9 MDA-MB-231, MCF7TamR In vivo Cas9n vector Immunofluorescence, A...…”

Strategies to overcome the main challenges of the use of CRISPR/Cas9 as a replacement for cancer therapy

“… [ 54 ] Breast cancer APOBEC3G MCF10A and HCC1806 - Knockout lipofection multiple clones evaluated for APOBEC3G gene knockout success. [ 55 ] Breast cancer CDK4, SRPK1, DNMT1 MCF10A, HEK 293T and GP2-293 Mice Knockout lentiviruses Transcriptional epistasis influences around 50% of differentially expressed genes in cancer cells. [ 56 ] Breast cancer CDH1 MCF-7 Rats Knockout Plasmid Transfection It is possible to target cancer-related genes using any genome editing technique.…”