Genome-scale CRISPR screen identifies TMEM41B as a critical host factor required for coronavirus replication [Focused cell]

We use the Focused-library to identify key host factors facilitating TGEV infection in porcine cells. We generated a Focused cell collection (in the PK-15 background) as previously described and performed a focused loss-of-function genetic screen. Briefly, we conducted five rounds of TGEV challenge, employing untreated PK-15-Cas9 cells as a negative control to confirm the cell death caused by TGEV infection in each round. The surviving mutant cells were collected and used for subsequent TGEV challenge rounds. Finally, the sgRNA constructs in surviving mutant cells of third and fifth rounds were PCR amplified and deep sequenced.