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Engineering digitizer circuits for chemical and genetic screens in human cells
- 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.
- Subjects :
- MAPK/ERK pathway
Computer science
Science
General Physics and Astronomy
Computational biology
Article
General Biochemistry, Genetics and Molecular Biology
Small Molecule Libraries
Genes, Reporter
Humans
CRISPR
Promoter Regions, Genetic
Gene
Synthetic biology
Electronic circuit
Multidisciplinary
High-throughput screening
Functional genomics
Genomics
General Chemistry
High-Throughput Screening Assays
Scalable system
CRISPR-Cas Systems
Transcription Factors
Genetic screen
Subjects
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