An arrayed CRISPR library for individual, combinatorial and multiplexed gene knockout II. synthetic construct

We have combined a machine-learning approach with other strategies to optimize the efficiency of sgRNAs for CRISPR screens and have constructed a genome-wide, sequence-verified, arrayed CRISPR library. This incorporates expression strategies to facilitate multiplexed or combinatorial screening. By conducting parallel loss-of-function screens, we compare our approach to existing sgRNA design and expression strategies. Overall design: High throughput sequencing was performed to identify the sgRNA representation in depletion screens at the initial (T0) or final (T6) timepoints, with 3 biological replicates per screen. We compared 5 different sgRNA libraries, with around 200 sgRNAs per library.