Overcoming CRISPR-Cas9 off-target prediction hurdles: A novel approach with ESB rebalancing strategy and CRISPR-MCA model
Fig 7
The Efficiency and Specificity-Based class rebalancing strategy encompasses two distinct phases: Initial mutation screening of gRNAs and subsequent specificity assessment of high-efficiency mutants.
Initially, each nucleotide within a gRNA is substituted with the three alternative nucleotides, following which the targeting efficiencies of these mutated gRNAs are evaluated. Mutants demonstrating superior efficiency compared to the original gRNA are selected for further analysis. The second phase involves identifying potential off-target sites across the genome using Cas-Offinder, succeeded by calculating the specificity of mutant gRNAs’ interaction with the target DNA sequence. Mutants that exhibit specificity surpassing that of the original sequence are then adopted as enhanced positive samples for subsequent training purposes.