A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens
Abstract Mapping genetic interactions is essential for determining gene function and defining novel biological pathways. We report a simple to use CRISPR interference (CRISPRi) based platform, compatible with Fluorescence Activated Cell Sorting (FACS)-based reporter screens, to query...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
BioMed Central
2023
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| Online Access: | https://hdl.handle.net/1721.1/152913 |
| _version_ | 1826193654289530880 |
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| author | Guna, Alina Page, Katharine R. Replogle, Joseph M. Esantsi, Theodore K. Wang, Maxine L. Weissman, Jonathan S. Voorhees, Rebecca M. |
| author2 | Whitehead Institute for Biomedical Research |
| author_facet | Whitehead Institute for Biomedical Research Guna, Alina Page, Katharine R. Replogle, Joseph M. Esantsi, Theodore K. Wang, Maxine L. Weissman, Jonathan S. Voorhees, Rebecca M. |
| author_sort | Guna, Alina |
| collection | MIT |
| description | Abstract
Mapping genetic interactions is essential for determining gene function and defining novel biological pathways. We report a simple to use CRISPR interference (CRISPRi) based platform, compatible with Fluorescence Activated Cell Sorting (FACS)-based reporter screens, to query epistatic relationships at scale. This is enabled by a flexible dual-sgRNA library design that allows for the simultaneous delivery and selection of a fixed sgRNA and a second randomized guide, comprised of a genome-wide library, with a single transduction. We use this approach to identify epistatic relationships for a defined biological pathway, showing both increased sensitivity and specificity than traditional growth screening approaches. |
| first_indexed | 2024-09-23T09:42:35Z |
| format | Article |
| id | mit-1721.1/152913 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T09:42:35Z |
| publishDate | 2023 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | mit-1721.1/1529132024-01-24T18:54:41Z A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens Guna, Alina Page, Katharine R. Replogle, Joseph M. Esantsi, Theodore K. Wang, Maxine L. Weissman, Jonathan S. Voorhees, Rebecca M. Whitehead Institute for Biomedical Research Howard Hughes Medical Institute Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Abstract Mapping genetic interactions is essential for determining gene function and defining novel biological pathways. We report a simple to use CRISPR interference (CRISPRi) based platform, compatible with Fluorescence Activated Cell Sorting (FACS)-based reporter screens, to query epistatic relationships at scale. This is enabled by a flexible dual-sgRNA library design that allows for the simultaneous delivery and selection of a fixed sgRNA and a second randomized guide, comprised of a genome-wide library, with a single transduction. We use this approach to identify epistatic relationships for a defined biological pathway, showing both increased sensitivity and specificity than traditional growth screening approaches. 2023-11-06T18:35:15Z 2023-11-06T18:35:15Z 2023-10-30 2023-11-05T04:12:18Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/152913 BMC Genomics. 2023 Oct 30;24(1):651 PUBLISHER_CC en https://doi.org/10.1186/s12864-023-09754-y Creative Commons Attribution https://creativecommons.org/licenses/by/4.0/ The Author(s) application/pdf BioMed Central BioMed Central |
| spellingShingle | Guna, Alina Page, Katharine R. Replogle, Joseph M. Esantsi, Theodore K. Wang, Maxine L. Weissman, Jonathan S. Voorhees, Rebecca M. A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title | A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title_full | A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title_fullStr | A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title_full_unstemmed | A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title_short | A dual sgRNA library design to probe genetic modifiers using genome-wide CRISPRi screens |
| title_sort | dual sgrna library design to probe genetic modifiers using genome wide crispri screens |
| url | https://hdl.handle.net/1721.1/152913 |
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