Wong, Nicole N., Frias, Elizabeth, Sigoillot, Frederic, Letendre, Justin, Hild, Marc and Wong, Wilson W. (2021) Engineering digitizer circuits for chemical and genetic screens in human cells. Nature communications, 12 (1). p. 6150. ISSN 2041-1723

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.

Item Type:	Article
Date Deposited:	30 Nov 2021 00:45
Last Modified:	30 Nov 2021 00:45
URI:	https://oak.novartis.com/id/eprint/44201