CRISPR screening core facility
The CRISPR screening facility holds and maintains gold-standard, pooled sgRNA libraries but can also generate bespoke, custom design libraries for pooled screening. Pooled CRISPR screening with cell viability readout is the primary screen format supported by the CRISPR facility, but other formats are also possible. During pooled screening, genomic perturbations (such as knock-outs) are generated simultaneously on an up to genome-wide level, one edit per cell, in large tissue culture vessels. Delivery of screen reagents is facilitated by the use of lentivirus, which the facility can also provide as validated high-titer stock.
Essential for successful screening is the evaluation and optimization of the assay system. Therefore the CRISPR facility can assist in the following steps: optimization of lentiviral transduction conditions, determination of antibiotic selection conditions and evaluation of screen treatments.
For the analysis of a pooled CRISPR screen with viability readout, the frequency of sgRNA barcodes (as a proxy for genotype abundance) is measured in screen endpoint samples by next-generation sequencing. Processing of samples for NGS and data analysis using published tools is also supported by the facility.