Genome-wide CRISPR guide RNA design and specificity analysis with GuideScan2.
| Publication Type | Academic Article |
| Authors | Schmidt H, Zhang M, Chakarov D, Bansal V, Mourelatos H, Sánchez-Rivera F, Lowe S, Ventura A, Leslie C, Pritykin Y |
| Journal | Genome Biol |
| Volume | 26 |
| Issue | 1 |
| Pagination | 41 |
| Date Published | 02/26/2025 |
| ISSN | 1474-760X |
| Keywords | RNA, Guide, CRISPR-Cas Systems, CRISPR-Cas Systems, Software |
| Abstract | 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. |
| DOI | 10.1186/s13059-025-03488-8 |
| PubMed ID | 40011959 |
| PubMed Central ID | PMC11863968 |