Development of CRISPR knockout screening in mosquito cell lines

Insect cell culture has been a valuable tool to dissect conserved pathways that are simpler in insects than mammals as well as insect-specific biology: interactions with insect hormones, pathogens, toxins, and drugs. Pooled format screens such as CRISPR screens had been unavailable in insect cell-lines due to an inability to conduct massively parallel delivery of large CRISPR guide libraries. In this poster, we share our results on developing CRISPR screening in Drosophila (fruit fly) cells and show a recently published use case. In unpublished work, we have now adapted a cell-line from Anopheles gambiae (malaria mosquito) and validate the approach using pilot gene-by- drug screens. We also describe an online resource for preparing CRISPR libraries for mosquito cell-lines.