Figure 1: Whole genome screening strategy for entry of SARS-CoV-2-S, VSVG-mNG and VSVG-CRE pseudoviruses. A) ACE2 293T or ACE2-293T-GFPflox cells were transduced with the Brunello sgRNA genome wide CRISPR library. These cells were selected with puromycin to kill off cells that did not integrate the sgRNA cassette. 5 days following selection, these cells were infected with SARS-CoV-2 Spike, VSV-G or VSV-G-Cre pseudoviruses and allowed 48h for reporter expression. The cells were sorted using FACS, genomic DNA was extracted, sgRNA sequences were amplified by PCR, sequenced by next generation sequencing and sgRNA abundance in the sample was quantified by MAGeCK. B) Representation of Cre-LoxP system in ACE2-293T-GFPflox cells. The cells express dsRed and after excision of dsRed-stop by Cre recombinase, the cells start to express EGFP.