Comparative analyses of the transcriptome of human non-small lung cancer cell lines (NSCLC) naturally expressing complement factor I (FI) and their CRISPR/Cas9 -edited clones with FI knock- out
Three NSCLC cell lines naturally expressing FI: H2087, H226, and H358 were genome-edited byCRISPR/Cas9 technology. Lentiviral constructs containing guide RNAs targeting FI (Forward:CACCGATATGAACATTGTCTTATCT, Reverse: AAACAGATAAGACAATGTTCATATC) were transfected intoHEK293T cells with the packaging enzymes psPAX2, pVSVg, pAdVAntage using polyethyleneimine(PEI; Polyscience) for virus production. Filtered viral supernatants were used for the transduction ofNSCLC cells by spinoculation in the presence of 8 mg/mL protamine sulfate. After transduction, cellswere selected based on the expression of GFP. Then, single cell-derived clonal lines were generated,subcultured and harvested for RNA isolation and sequencing.