Deep Sampling Of Grna In The Human Genome And Deep-learning-informed Prediction Of Grna Activities

This study designed a 740k gRNA-target pair library corresponding to 26 gRNAs per protein-coding gene and 12 gRNAs per non-coding gene. The library was a combination of the gRNAs that were newly designed for this study and other published libraries. The assessment cassette for each gRNA included the gRNA itself and its 63-nucleotides putative genome target. Transduction was conducted at a multiplicity of infection (MOI) of 5 to achieve high cell-to-library coverage while generating a cell population that was manageable in size. The indel frequencies of all gRNAs were assessed by high-throughput sequencing (HTS) and the frequency spectra collectively showed that the indel rates were widely distributed.