Crispr screens in pools

Crispr screens in pools allow you to identify genes involved in a given cell function. All we need is a library encoding sgRNAs for all or a subset of the transcriptome, usually in a lentiviral vector, and a cell assay that acts as a selection step. Once the cells have been transduced with the library, they undergo the selection step (for instance, exposure to a drug) and then we amplify and sequence the integrated sgRNA encoding regions from the cell pools before and after the selection. From the high-throughput sequencing data we can compare the frequencies of each individual sgRNA encoding region before and after the selection step. Any statistically significant difference (corrected for multiple comparisons) is likely  to identify a gene that encodes a protein or RNA important for the selection step, by increasing the sensitivity or resistance of the cell.

Our Crispr screens start at 25k euros (turnaround time is 7 months) and include:

1. Amplification of the library in commercially electrocompetent E. coli to up to at least 2-5 mg of DNA, keeping the original complexity (at least 200x the size of the library). The library must be provided by the client. Addgene is a good source of pooled libraries and we can help you choosing the most suitable for your project.
2. Optimization of transduction and freezing/thawing for maximum viability. 
3. Transduction of the cells with the library and the selection step.
4. Amplification and high-throughput sequencing of the gRNA encoding regions, following the best recommendations in the field.
5. Analysis of the data to identify the most interesting candidate genes.
6. Delivery of the originally transduced cells (at least 100 millions).