CRISPR-Cas9 Engineered Haploid Cell Lines

Human-engineered haploid cells (eHAP cells; Horizon Discovery), eHAP cells with both huANP32A and huANP32B (eHAP dKO) ablated via CRISPR-Cas9, as previously described (14 (link)), or eHAP cells with huANP32A, huANP32B, and huANP32E ablated via CRISPR-Cas9 (eHAP tKO) (gift from Ecco Staller and Ervin Fodor) were maintained in Iscove modified Dulbecco medium (Thermo Fisher) supplemented with 10% fetal bovine serum (FBS; Labtech), 1% penicillin-streptomycin (pen-strep; Gibco), and 1% nonessential amino acids (NEAA; Gibco). Human embryonic kidney (293Ts; ATCC) and Madin-Darby canine kidney (ATCC) cells were maintained in Dulbecco modified Eagle medium supplemented with 10% FBS, 1% pen-strep, and 1% NEAA. When used for infection, 293T cells were cultured on poly-l-lysine-coated plates to aid adherence. All cells were maintained at 37°C and 5% CO₂.