TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways
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Publication Date
2022-01-20Journal Title
Molecular Cell
ISSN
1097-2765
Publisher
Cell Press
Type
Article
This Version
AM
Metadata
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Tchasovnikarova, I., Kingston, R., Marr, S., & Damle, M. (2022). TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways. Molecular Cell https://doi.org/10.17863/CAM.79100
Abstract
Genetically-encoded biosensors are powerful tools to monitor cellular behavior, but the difficulty in generating appropriate reporters for chromatin factors hampers our ability to dissect epigenetic pathways. Here we present TRACE, a high-throughput, genome-wide technique to generate fluorescent human reporter cell lines responsive to manipulation of epigenetic factors. By profiling GFP expression from a large pool of individually barcoded lentiviral integrants in the presence and absence of a perturbation, we identify reporters responsive to pharmacological inhibition of the histone lysine demethylase LSD1 and genetic ablation of the PRC2 subunit SUZ12. Furthermore, by manipulating the HIV-1 host factor LEDGF through targeted deletion or fusion to chromatin reader domains, we alter lentiviral integration site preferences, thus broadening the types of chromatin examined by TRACE. The phenotypic reporters generated through TRACE will allow the genetic interrogation of a broad range of epigenetic pathways, furthering our mechanistic understanding of chromatin biology.
Sponsorship
Damon Runyon Cancer Research Foundation
NIH
Funder references
Damon Runyon Cancer Research Foundation (42-20)
Embargo Lift Date
2022-12-27
Identifiers
This record's DOI: https://doi.org/10.17863/CAM.79100
This record's URL: https://www.repository.cam.ac.uk/handle/1810/331649
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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