A synthetic gene circuit for imaging-free detection of signaling pulses.

TitleA synthetic gene circuit for imaging-free detection of signaling pulses.
Publication TypeJournal Article
Year of Publication2022
AuthorsRavindran, PT, McFann, S, Thornton, RH, Toettcher, JE
JournalCell Syst
Volume13
Issue2
Pagination131-142.e13
Date Published2022 Feb 16
ISSN2405-4720
KeywordsGene Expression Regulation, Gene Regulatory Networks, Genes, Synthetic, MAP Kinase Signaling System, Signal Transduction
Abstract

<p>Cells employ intracellular signaling pathways to sense and respond to changes in their external environment. In recent years, live-cell biosensors have revealed complex pulsatile dynamics in many pathways, but studies of these signaling dynamics are limited by the necessity of live-cell imaging at high spatiotemporal resolution. Here, we describe an approach to infer pulsatile signaling dynamics from a single measurement in fixed cells using a pulse-detecting gene circuit. We computationally screened for circuits with the capability to selectively detect signaling pulses, revealing an incoherent feedforward topology that robustly performs this computation. We implemented the motif experimentally for the Erk signaling pathway using a single engineered transcription factor and fluorescent protein reporter. Our "recorder of Erk activity dynamics" (READer) responds sensitively to spontaneous and stimulus-driven Erk pulses. READer circuits open the door to permanently labeling transient, dynamic cell populations to elucidate the mechanistic underpinnings and biological consequences of signaling dynamics.</p>

DOI10.1016/j.cels.2021.10.002
Alternate JournalCell Syst
PubMed ID34739875
PubMed Central IDPMC8857027
Grant ListDP2 EB024247 / EB / NIBIB NIH HHS / United States
T32 GM007388 / GM / NIGMS NIH HHS / United States