TY - JOUR
AU - Yin, Jiang-An
AU - Frick, Lukas
AU - Scheidmann, Manuel C
AU - Liu, Tingting
AU - Trevisan, Chiara
AU - Dhingra, Ashutosh
AU - Spinelli, Anna
AU - Wu, Yancheng
AU - Yao, Longping
AU - Vena, Dalila Laura
AU - Knapp, Britta
AU - Guo, Jingjing
AU - De Cecco, Elena
AU - Ging, Kathi
AU - Armani, Andrea
AU - Oakeley, Edward J
AU - Nigsch, Florian
AU - Jenzer, Joel
AU - Haegele, Jasmin
AU - Pikusa, Michal
AU - Täger, Joachim
AU - Rodriguez-Nieto, Salvador
AU - Bouris, Vangelis
AU - Ribeiro, Rafaela
AU - Baroni, Federico
AU - Bedi, Manmeet Sakshi
AU - Berry, Scott
AU - Losa, Marco
AU - Hornemann, Simone
AU - Kampmann, Martin
AU - Pelkmans, Lucas
AU - Hoepfner, Dominic
AU - Heutink, Peter
AU - Aguzzi, Adriano
TI - Arrayed CRISPR libraries for the genome-wide activation, deletion and silencing of human protein-coding genes.
JO - Nature biomedical engineering
VL - 9
IS - 1
SN - 2157-846X
CY - Tokyo
PB - Nature Research
M1 - DZNE-2025-00230
SP - 127 - 148
PY - 2025
AB - 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
KW - Humans
KW - Gene Silencing
KW - CRISPR-Cas Systems: genetics
KW - Genome, Human: genetics
KW - Gene Library
KW - RNA, Guide, CRISPR-Cas Systems: genetics
KW - Plasmids: genetics
KW - Gene Deletion
KW - HEK293 Cells
KW - Clustered Regularly Interspaced Short Palindromic Repeats: genetics
KW - Transcription Factors: genetics
KW - Transcription Factors: metabolism
KW - RNA, Guide, CRISPR-Cas Systems (NLM Chemicals)
KW - Transcription Factors (NLM Chemicals)
LB - PUB:(DE-HGF)16
C6 - pmid:39633028
DO - DOI:10.1038/s41551-024-01278-4
UR - https://pub.dzne.de/record/276158
ER -