Figure 1.

rh32.33 Entry Screen

(A) Huh7 AAVR KO cells undergo lentiCRISPR mutagenesis using a two-vector lentiviral GeCKO system. sgRNA library contains six sgRNAs per gene, targeting the entire human genome and 1,000 miRNAs to generate a cell line knockout library. Cells are subjected to multiple rounds of high MOI transduction and sgRNA deep sequencing to determine gene deletions enriched in GFP-negative cells to identify genes required for rh32.33 entry. (B) Entry screen hits from second round of GFP transduction. The x axis shows individual genes within the GeCKO library, grouped by gene ontology analysis. The y axis shows significance of hit based on RRA analysis. The bubble diameter indicates the number of individual sgRNAs per gene enriched in the selected population, relative to unselected control.