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bioRxiv · 10.1101/2022.05.02.490368

Genome-wide CRISPR guide RNA design and specificity analysis with GuideScan2

Schmidt, H.·Zhang, M.·Mourelatos, H.·Sanchez-Rivera, F. J.·Lowe, S. W.·Ventura, A.·Leslie, C. S.·Pritykin, Y.
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Abstract

We present GuideScan2 for memory-efficient, parallelizable construction of high-specificity CRISPR guide RNA (gRNA) databases and user-friendly gRNA/library design in custom genomes. GuideScan2 analysis identified widespread confounding effects of low-specificity gRNAs in published CRISPR knockout, interference and activation screens and enabled construction of a ready-to-use gRNA library that reduced off-target effects in a novel gene essentiality screen. GuideScan2 also enabled the design and experimental validation of allele-specific gRNAs in a hybrid mouse genome.

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Schmidt, H., Zhang, M., Mourelatos, H., Sanchez-Rivera, F. J., Lowe, S. W., Ventura, A., Leslie, C. S., Pritykin, Y.. 2022-05-03. Genome-wide CRISPR guide RNA design and specificity analysis with GuideScan2. https://doi.org/10.1101/2022.05.02.490368

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