%0 Journal Article
%A Yin, Jiang-An
%A Frick, Lukas
%A Scheidmann, Manuel C
%A Liu, Tingting
%A Trevisan, Chiara
%A Dhingra, Ashutosh
%A Spinelli, Anna
%A Wu, Yancheng
%A Yao, Longping
%A Vena, Dalila Laura
%A Knapp, Britta
%A Guo, Jingjing
%A De Cecco, Elena
%A Ging, Kathi
%A Armani, Andrea
%A Oakeley, Edward J
%A Nigsch, Florian
%A Jenzer, Joel
%A Haegele, Jasmin
%A Pikusa, Michal
%A Täger, Joachim
%A Rodriguez-Nieto, Salvador
%A Bouris, Vangelis
%A Ribeiro, Rafaela
%A Baroni, Federico
%A Bedi, Manmeet Sakshi
%A Berry, Scott
%A Losa, Marco
%A Hornemann, Simone
%A Kampmann, Martin
%A Pelkmans, Lucas
%A Hoepfner, Dominic
%A Heutink, Peter
%A Aguzzi, Adriano
%T Arrayed CRISPR libraries for the genome-wide activation, deletion and silencing of human protein-coding genes.
%J Nature biomedical engineering
%V 9
%N 1
%@ 2157-846X
%C Tokyo
%I Nature Research
%M DZNE-2025-00230
%P 127 - 148
%D 2025
%X 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
%K Humans
%K Gene Silencing
%K CRISPR-Cas Systems: genetics
%K Genome, Human: genetics
%K Gene Library
%K RNA, Guide, CRISPR-Cas Systems: genetics
%K Plasmids: genetics
%K Gene Deletion
%K HEK293 Cells
%K Clustered Regularly Interspaced Short Palindromic Repeats: genetics
%K Transcription Factors: genetics
%K Transcription Factors: metabolism
%K RNA, Guide, CRISPR-Cas Systems (NLM Chemicals)
%K Transcription Factors (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:39633028
%R 10.1038/s41551-024-01278-4
%U https://pub.dzne.de/record/276158