@article {1471848,
	title = {Pooled CRISPR Screens in Drosophila Cells},
	journal = {Current Protocols in Molecular Biology},
	volume = {129},
	year = {2019},
	abstract = {High-throughput screens in Drosophila melanogaster cell lines have led to discovery of conserved gene functions related to signal transduction, host pathogen interactions, ion transport, and more. CRISPR/Cas9 technology has opened the door to new types of large-scale cell-based screens. Whereas array format screens require liquid handling automation and assay miniaturization, pooled-format screens, in which reagents are introduced at random and in bulk, can be done in a standard lab setting. We provide a detailed protocol for conducting and evaluating genome-wide CRISPR single guide RNA (sgRNA) pooled screens in Drosophila S2R+ cultured cells. Specifically, we provide step-by-step instructions for library design and production, optimization of cytotoxin-based selection assays, genome-scale screening, and data analysis. This type of project takes ~3 months to complete. Results can be used in follow-up studies performed in vivo in Drosophila, mammalian cells, and/or other systems.},
	author = {Viswanatha, R and Brathwaite, R and Hu, Y. and Li, Z. and Rodiger, J and Merckaert, P and Chung, V and Mohr, SE and Perrimon, N}
}
