Figure 1.

Genome-wide CRISPR screen for genetic regulators of phagocytosis of bacteria. THP-1 iCas9 cells were transduced at a coverage of 2919× with a genome-wide pooled sgRNA library containing 6 sgRNAs per gene and 500 non-targeting controls. Transduced cells were enriched by MACS and Cas9 was induced with dox on two consecutive days. 14 days after transduction, S. aureus particles labeled with pHrodo red was added to the cells. After 60 min, cells were sorted by FACS according to their fluorescence intensity in phagocytic active and inactive populations. The integrated sgRNAs were amplified and sequenced with NextSeq 550. Sequencing data was analyzed with MAGeCK-VISPR.