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<p>We present GuideScan2 for memory-efficient, parallelizable construction of high-specificity CRISPR guide RNA (gRNA) databases and user-friendly design and analysis of individual gRNAs and gRNA libraries for targeting coding and non-coding regions in custom genomes. GuideScan2 analysis identifies widespread confounding effects of low-specificity gRNAs in published CRISPR screens and enables construction of a gRNA library that reduces off-target effects in a gene essentiality screen. GuideScan2 also enables the design and experimental validation of allele-specific gRNAs in a hybrid mouse genome. GuideScan2 will facilitate CRISPR experiments across a wide range of applications.</p>

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

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年2月26日

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  • Henri Schmidt,
  • Minsi Zhang,
  • Dimitar Chakarov,
  • Vineet Bansal,
  • Haralambos Mourelatos,
  • Francisco J. Sánchez-Rivera,
  • Scott W. Lowe,
  • Andrea Ventura,
  • Christina S. Leslie,
  • Yuri Pritykin

摘要

We present GuideScan2 for memory-efficient, parallelizable construction of high-specificity CRISPR guide RNA (gRNA) databases and user-friendly design and analysis of individual gRNAs and gRNA libraries for targeting coding and non-coding regions in custom genomes. GuideScan2 analysis identifies widespread confounding effects of low-specificity gRNAs in published CRISPR screens and enables construction of a gRNA library that reduces off-target effects in a gene essentiality screen. GuideScan2 also enables the design and experimental validation of allele-specific gRNAs in a hybrid mouse genome. GuideScan2 will facilitate CRISPR experiments across a wide range of applications.