Does every sgRNA in the library work?

It is impossible to test all 76,456 sgRNAs functionally. However, the target sequences cloned into the Guide-it Genome-Wide sgRNA Library are the same as those described by Doench et al. in 2016. In this paper, the authors created and modified their Brunello algorithm based on real screening and activity data from positive and negative screens. At Takara Bio we verified the editing activity of a selected number of clones chosen at random and confirmed that all worked well.
ReferenceDoench, J. G. et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR Cas9. Nat. Biotechnol. 34, 184–191 (2016).

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