Submitted by est4003 on August 20, 2025 - 11:20am
| Title | Genome-wide CRISPR guide RNA design and specificity analysis with GuideScan2. |
| Publication Type | Journal Article |
| Year of Publication | 2025 |
| Authors | Schmidt H, Zhang M, Chakarov D, Bansal V, Mourelatos H, Sánchez-Rivera FJ, Lowe SW, Ventura A, Leslie CS, Pritykin Y |
| Journal | Genome Biol |
| Volume | 26 |
| Issue | 1 |
| Pagination | 41 |
| Date Published | 2025 Feb 26 |
| ISSN | 1474-760X |
| Keywords | Animals, CRISPR-Cas Systems, Genome, Mice, RNA, Guide, 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 |
| Alternate Journal | Genome Biol |
| PubMed ID | 40011959 |
| PubMed Central ID | PMC11863968 |
| Grant List | T32 CA160001 / CA / NCI NIH HHS / United States DP2 AI171161 / NH / NIH HHS / United States P01 CA129243 / NH / NIH HHS / United States U01 HG012103 / HG / NHGRI NIH HHS / United States P30 CA008748 / CA / NCI NIH HHS / United States 5T32CA160001 / / MSKCC TROT program / DP2 AI171161 / AI / NIAID NIH HHS / United States U01 HG009395 / NH / NIH HHS / United States CAREER 2238831 / / National Science Foundation / |