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@ARTICLE{Yin:276158,
author = {Yin, Jiang-An and Frick, Lukas and Scheidmann, Manuel C and
Liu, Tingting and Trevisan, Chiara and Dhingra, Ashutosh and
Spinelli, Anna and Wu, Yancheng and Yao, Longping and Vena,
Dalila Laura and Knapp, Britta and Guo, Jingjing and De
Cecco, Elena and Ging, Kathi and Armani, Andrea and Oakeley,
Edward J and Nigsch, Florian and Jenzer, Joel and Haegele,
Jasmin and Pikusa, Michal and Täger, Joachim and
Rodriguez-Nieto, Salvador and Bouris, Vangelis and Ribeiro,
Rafaela and Baroni, Federico and Bedi, Manmeet Sakshi and
Berry, Scott and Losa, Marco and Hornemann, Simone and
Kampmann, Martin and Pelkmans, Lucas and Hoepfner, Dominic
and Heutink, Peter and Aguzzi, Adriano},
title = {{A}rrayed {CRISPR} libraries for the genome-wide
activation, deletion and silencing of human protein-coding
genes.},
journal = {Nature biomedical engineering},
volume = {9},
number = {1},
issn = {2157-846X},
address = {Tokyo},
publisher = {Nature Research},
reportid = {DZNE-2025-00230},
pages = {127 - 148},
year = {2025},
abstract = {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.},
keywords = {Humans / Gene Silencing / CRISPR-Cas Systems: genetics /
Genome, Human: genetics / Gene Library / RNA, Guide,
CRISPR-Cas Systems: genetics / Plasmids: genetics / Gene
Deletion / HEK293 Cells / Clustered Regularly Interspaced
Short Palindromic Repeats: genetics / Transcription Factors:
genetics / Transcription Factors: metabolism / RNA, Guide,
CRISPR-Cas Systems (NLM Chemicals) / Transcription Factors
(NLM Chemicals)},
cin = {AG Heutink / AG Täger},
ddc = {610},
cid = {I:(DE-2719)1210002 / I:(DE-2719)5000002},
pnm = {354 - Disease Prevention and Healthy Aging (POF4-354) / 899
- ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-354 / G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39633028},
doi = {10.1038/s41551-024-01278-4},
url = {https://pub.dzne.de/record/276158},
}