We wish to perform large scale genomic CRISPR screens like the DepMAp project for drug resistance studies. Most negative screens suggest 300-1000X coverage of a library, and the library we will use has 100,000 gRNAs in one tube. Just wondering how the BROAD can routinely infect all kinds of cell lines at efficiency around 30% to generate at least 30 million infected cells initially before cell amplification, which assumes that 90 million cells are in the original infection step. This seems too many cells for spin inoculation, and many cell lines have very poor infection efficiency even with reverse infection… What’s the experimental method or secret to make this feasible? Is this still done as described in original papers or has this been refined/updated. Thanks
Hi!
The Broad Genetic Perturbation Platform is the platform used by DepMap for these screens. The GPP has a protocols page that can be used as a reference for this information.
Thanks,
The DepMap Team
