Engineering digitizer circuits for chemical and genetic screens in human cells

Nicole M. Wong, Elizabeth Frias, Frederic D. Sigoillot, Justin H. Letendre, Marc Hild () and Wilson W. Wong ()
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Nicole M. Wong: Boston University
Elizabeth Frias: Novartis Institutes for BioMedical Research
Frederic D. Sigoillot: Novartis Institutes for BioMedical Research
Justin H. Letendre: Boston University
Marc Hild: Novartis Institutes for BioMedical Research
Wilson W. Wong: Boston University

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-021-26359-9

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