Abstract: A specific target site obtained at the PAS (Per-ARNT-Sim) domain of human HIF-1α was used to construct a Cas9-gRNA-HIF-1α knockout plasmid. After transfected it into KGN cells, 21 monoclonal cell lines were selected by puromycin, and then determined by enzyme digestion, sequencing analysis and downstream gene verification, and finally obtained a specifically knockout HIF-1α homozygous cells (H20). Collectively, the specific knockout site used in this study can be efficiently target against HIF-1α, which will provide a reference for the specific knockout of HIF-1α in other cells through CRISPR/Cas9 system.

Key words: HIF-1α, CRISPR/Cas9, gene knockout