Compact CRISPR genetic screens enabled by improved guide RNA library cloning
Abstract CRISPR genome editing approaches theoretically enable researchers to define the function of each human gene in specific cell types, but challenges remain to efficiently perform genetic perturbations in relevant models. In this work, we develop a library cloning protocol that increases sgRNA...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-01-01
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| Series: | Genome Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13059-023-03132-3 |
| _version_ | 1797349927475478528 |
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| author | Seok-Jin Heo Lauren D. Enriquez Scot Federman Amy Y. Chang Rachel Mace Kaivalya Shevade Phuong Nguyen Adam J. Litterman Shawn Shafer Laralynne Przybyla Eric D. Chow |
| author_facet | Seok-Jin Heo Lauren D. Enriquez Scot Federman Amy Y. Chang Rachel Mace Kaivalya Shevade Phuong Nguyen Adam J. Litterman Shawn Shafer Laralynne Przybyla Eric D. Chow |
| author_sort | Seok-Jin Heo |
| collection | DOAJ |
| description | Abstract CRISPR genome editing approaches theoretically enable researchers to define the function of each human gene in specific cell types, but challenges remain to efficiently perform genetic perturbations in relevant models. In this work, we develop a library cloning protocol that increases sgRNA uniformity and greatly reduces bias in existing genome-wide libraries. We demonstrate that our libraries can achieve equivalent or better statistical power compared to previously reported screens using an order of magnitude fewer cells. This improved cloning protocol enables genome-scale CRISPR screens in technically challenging cell models and screen formats. |
| first_indexed | 2024-03-08T12:37:27Z |
| format | Article |
| id | doaj.art-5f026eebf6b34406bec636b6eb2f851a |
| institution | Directory Open Access Journal |
| issn | 1474-760X |
| language | English |
| last_indexed | 2024-03-08T12:37:27Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Genome Biology |
| spelling | doaj.art-5f026eebf6b34406bec636b6eb2f851a2024-01-21T12:23:35ZengBMCGenome Biology1474-760X2024-01-0125112010.1186/s13059-023-03132-3Compact CRISPR genetic screens enabled by improved guide RNA library cloningSeok-Jin Heo0Lauren D. Enriquez1Scot Federman2Amy Y. Chang3Rachel Mace4Kaivalya Shevade5Phuong Nguyen6Adam J. Litterman7Shawn Shafer8Laralynne Przybyla9Eric D. Chow10Laboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchLaboratory for Genomics ResearchAbstract CRISPR genome editing approaches theoretically enable researchers to define the function of each human gene in specific cell types, but challenges remain to efficiently perform genetic perturbations in relevant models. In this work, we develop a library cloning protocol that increases sgRNA uniformity and greatly reduces bias in existing genome-wide libraries. We demonstrate that our libraries can achieve equivalent or better statistical power compared to previously reported screens using an order of magnitude fewer cells. This improved cloning protocol enables genome-scale CRISPR screens in technically challenging cell models and screen formats.https://doi.org/10.1186/s13059-023-03132-3CRISPR screeningLibrary cloningGuide libraryiPSC-derived CRISPR screeningPrimary cell CRISPR screening |
| spellingShingle | Seok-Jin Heo Lauren D. Enriquez Scot Federman Amy Y. Chang Rachel Mace Kaivalya Shevade Phuong Nguyen Adam J. Litterman Shawn Shafer Laralynne Przybyla Eric D. Chow Compact CRISPR genetic screens enabled by improved guide RNA library cloning Genome Biology CRISPR screening Library cloning Guide library iPSC-derived CRISPR screening Primary cell CRISPR screening |
| title | Compact CRISPR genetic screens enabled by improved guide RNA library cloning |
| title_full | Compact CRISPR genetic screens enabled by improved guide RNA library cloning |
| title_fullStr | Compact CRISPR genetic screens enabled by improved guide RNA library cloning |
| title_full_unstemmed | Compact CRISPR genetic screens enabled by improved guide RNA library cloning |
| title_short | Compact CRISPR genetic screens enabled by improved guide RNA library cloning |
| title_sort | compact crispr genetic screens enabled by improved guide rna library cloning |
| topic | CRISPR screening Library cloning Guide library iPSC-derived CRISPR screening Primary cell CRISPR screening |
| url | https://doi.org/10.1186/s13059-023-03132-3 |
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