@article{oai:tsukuba.repo.nii.ac.jp:00041671,
 author = {渡邊, 幸秀 and 加藤, 光保 and Wardhani, Bantari W.K. and Puteri, Meidi U. and Watanabe, Yukihide and Louisa, Melva and Setiabudy, Rianto and Kato, Mitsuyasu},
 issue = {1},
 journal = {Medical journal of Indonesia},
 month = {May},
 note = {Background:
 Clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) is a powerful genome editing technique. It consists of RNA-guided DNA endonuclease Cas9 and single guide RNA (gRNA). By combining their expressions, high efficiency cleavage of the target gene can be achieved, leading to the formation of DNA double-strand break (DSB) at the genomic locus of interest which will be repaired via NHEJ (non-homologous end joining) or HDR (homology-directed repair) and mediate DNA alteration. We aimed to apply the CRISPR/Cas9 technique to knock-out the transmembrane prostate androgen-induced protein (TMEPAI) gene in the triple negative breast cancer cell line.
Methods:
 Designed gRNA which targets the TMEPAI gene was synthesized, annealed, and cloned into gRNA expression vector. It was co-transfected into the TNBC cell line using polyethylenimine (PEI) together with Cas9-GFP and puromycin resistant gene vector. At 24-hours post-transfection, cells were selected by puromycin for 3 days before they were cloned. Selected knock-out clones were subsequently checked on their protein levels by western blotting.
Results:
CRISPR/Cas9, a genome engineering technique successfully knocked-out TMEPAI in the Hs578T TNBC cell line. Sequencing shows a frameshift mutation in TMEPAI. Western blot shows the absence of TMEPAI band on Hs578T KO cells.
Conclusion: 
TMEPAI gene was deleted in the TNBC cell line using the genomic editing technique CRISPR/Cas9. The deletion was confirmed by genome and protein analysis.},
 pages = {14--18},
 title = {TMEPAI genome editing in triple negative breast cancer cells},
 volume = {26},
 year = {2017}
}