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Engineering digitizer circuits for chemical and genetic screens in human cells

Authors :
Nicole M. L. Wong
Elizabeth Frias
Wilson W. Wong
Justin H. Letendre
Frederic Sigoillot
Marc Hild
Source :
Nature Communications, Nature Communications, Vol 12, Iss 1, Pp 1-12 (2021)
Publication Year :
2021
Publisher :
Springer Science and Business Media LLC, 2021.

Abstract

Cell-based transcriptional reporters are invaluable in high-throughput compound and CRISPR screens for identifying compounds or genes that can impact a pathway of interest. However, many transcriptional reporters have weak activities and transient responses. This can result in overlooking therapeutic targets and compounds that are difficult to detect, necessitating the resource-consuming process of running multiple screens at various timepoints. Here, we present RADAR, a digitizer circuit for amplifying reporter activity and retaining memory of pathway activation. Reporting on the AP-1 pathway, our circuit identifies compounds with known activity against PKC-related pathways and shows an enhanced dynamic range with improved sensitivity compared to a classical reporter in compound screens. In the first genome-wide pooled CRISPR screen for the AP-1 pathway, RADAR identifies canonical genes from the MAPK and PKC pathways, as well as non-canonical regulators. Thus, our scalable system highlights the benefit and versatility of using genetic circuits in large-scale cell-based screening.<br />Cell-based transcriptional reporters are an invaluable part of highthroughput screening, but many such reporters have weak or transient responses. Here, the authors describe a digitizer circuit for amplifying reporter activity, increasing sensitivity, and retaining memory of pathway activation.

Details

ISSN :
20411723
Volume :
12
Database :
OpenAIRE
Journal :
Nature Communications
Accession number :
edsair.doi.dedup.....60f59203ae85a2d9b30704dd0b809ca8
Full Text :
https://doi.org/10.1038/s41467-021-26359-9