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.