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| 024 | 7 | _ | |a 10.1038/s41551-024-01278-4 |2 doi |
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| 037 | _ | _ | |a DZNE-2025-00230 |
| 041 | _ | _ | |a English |
| 082 | _ | _ | |a 610 |
| 100 | 1 | _ | |a Yin, Jiang-An |0 P:(DE-2719)9002756 |b 0 |
| 245 | _ | _ | |a Arrayed CRISPR libraries for the genome-wide activation, deletion and silencing of human protein-coding genes. |
| 260 | _ | _ | |a Tokyo |c 2025 |b Nature Research |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 520 | _ | _ | |a Arrayed CRISPR libraries extend the scope of gene-perturbation screens to non-selectable cell phenotypes. However, library generation requires assembling thousands of vectors expressing single-guide RNAs (sgRNAs). Here, by leveraging massively parallel plasmid-cloning methodology, we show that arrayed libraries can be constructed for the genome-wide ablation (19,936 plasmids) of human protein-coding genes and for their activation and epigenetic silencing (22,442 plasmids), with each plasmid encoding an array of four non-overlapping sgRNAs designed to tolerate most human DNA polymorphisms. The quadruple-sgRNA libraries yielded high perturbation efficacies in deletion (75-99%) and silencing (76-92%) experiments and substantial fold changes in activation experiments. Moreover, an arrayed activation screen of 1,634 human transcription factors uncovered 11 novel regulators of the cellular prion protein PrPC, screening with a pooled version of the ablation library led to the identification of 5 novel modifiers of autophagy that otherwise went undetected, and 'post-pooling' individually produced lentiviruses eliminated template-switching artefacts and enhanced the performance of pooled screens for epigenetic silencing. Quadruple-sgRNA arrayed libraries are a powerful and versatile resource for targeted genome-wide perturbations. |
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| 650 | _ | 7 | |a RNA, Guide, CRISPR-Cas Systems |2 NLM Chemicals |
| 650 | _ | 7 | |a Transcription Factors |2 NLM Chemicals |
| 650 | _ | 2 | |a Humans |2 MeSH |
| 650 | _ | 2 | |a Gene Silencing |2 MeSH |
| 650 | _ | 2 | |a CRISPR-Cas Systems: genetics |2 MeSH |
| 650 | _ | 2 | |a Genome, Human: genetics |2 MeSH |
| 650 | _ | 2 | |a Gene Library |2 MeSH |
| 650 | _ | 2 | |a RNA, Guide, CRISPR-Cas Systems: genetics |2 MeSH |
| 650 | _ | 2 | |a Plasmids: genetics |2 MeSH |
| 650 | _ | 2 | |a Gene Deletion |2 MeSH |
| 650 | _ | 2 | |a HEK293 Cells |2 MeSH |
| 650 | _ | 2 | |a Clustered Regularly Interspaced Short Palindromic Repeats: genetics |2 MeSH |
| 650 | _ | 2 | |a Transcription Factors: genetics |2 MeSH |
| 650 | _ | 2 | |a Transcription Factors: metabolism |2 MeSH |
| 700 | 1 | _ | |a Frick, Lukas |b 1 |
| 700 | 1 | _ | |a Scheidmann, Manuel C |b 2 |
| 700 | 1 | _ | |a Liu, Tingting |b 3 |
| 700 | 1 | _ | |a Trevisan, Chiara |0 0000-0002-3997-094X |b 4 |
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| 700 | 1 | _ | |a Spinelli, Anna |b 6 |
| 700 | 1 | _ | |a Wu, Yancheng |0 0000-0003-2809-4865 |b 7 |
| 700 | 1 | _ | |a Yao, Longping |b 8 |
| 700 | 1 | _ | |a Vena, Dalila Laura |0 0009-0007-6863-7349 |b 9 |
| 700 | 1 | _ | |a Knapp, Britta |b 10 |
| 700 | 1 | _ | |a Guo, Jingjing |b 11 |
| 700 | 1 | _ | |a De Cecco, Elena |b 12 |
| 700 | 1 | _ | |a Ging, Kathi |b 13 |
| 700 | 1 | _ | |a Armani, Andrea |0 0000-0002-4765-9748 |b 14 |
| 700 | 1 | _ | |a Oakeley, Edward J |0 0000-0001-6226-7470 |b 15 |
| 700 | 1 | _ | |a Nigsch, Florian |0 0000-0002-2919-8749 |b 16 |
| 700 | 1 | _ | |a Jenzer, Joel |b 17 |
| 700 | 1 | _ | |a Haegele, Jasmin |b 18 |
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| 700 | 1 | _ | |a Bouris, Vangelis |b 22 |
| 700 | 1 | _ | |a Ribeiro, Rafaela |b 23 |
| 700 | 1 | _ | |a Baroni, Federico |b 24 |
| 700 | 1 | _ | |a Bedi, Manmeet Sakshi |b 25 |
| 700 | 1 | _ | |a Berry, Scott |b 26 |
| 700 | 1 | _ | |a Losa, Marco |0 0000-0003-3428-418X |b 27 |
| 700 | 1 | _ | |a Hornemann, Simone |0 0000-0002-2674-9891 |b 28 |
| 700 | 1 | _ | |a Kampmann, Martin |b 29 |
| 700 | 1 | _ | |a Pelkmans, Lucas |b 30 |
| 700 | 1 | _ | |a Hoepfner, Dominic |b 31 |
| 700 | 1 | _ | |a Heutink, Peter |0 P:(DE-2719)2810728 |b 32 |
| 700 | 1 | _ | |a Aguzzi, Adriano |b 33 |
| 773 | _ | _ | |a 10.1038/s41551-024-01278-4 |g Vol. 9, no. 1, p. 127 - 148 |0 PERI:(DE-600)2878897-7 |n 1 |p 127 - 148 |t Nature biomedical engineering |v 9 |y 2025 |x 2157-846X |
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