TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways.
View / Open Files
Authors
Tchasovnikarova, Iva A
Marr, Sharon K
Damle, Manashree
Kingston, Robert E
Publication Date
2022-01-20Journal Title
Mol Cell
ISSN
1097-2765
Publisher
Elsevier BV
Type
Article
This Version
AM
Metadata
Show full item recordCitation
Tchasovnikarova, I. A., Marr, S. K., Damle, M., & Kingston, R. E. (2022). TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways.. Mol Cell https://doi.org/10.1016/j.molcel.2021.11.035
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 (transgene reporters across chromatin environments), 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.
Keywords
LEDGF, PRC2, Polycomb, SUZ12, TRACE, TRIP, chromatin, epigenetics, fluorescent reporter, lentiviral integration, Adaptor Proteins, Signal Transducing, Biosensing Techniques, Chromatin Assembly and Disassembly, Epigenesis, Genetic, Epigenome, Genes, Reporter, Genetic Vectors, Green Fluorescent Proteins, HEK293 Cells, HeLa Cells, Histone Demethylases, Humans, Lentivirus, Neoplasm Proteins, THP-1 Cells, Transcription Factors
Sponsorship
Damon Runyon Cancer Research Foundation
NIH
Funder references
Damon Runyon Cancer Research Foundation (42-20)
Identifiers
External DOI: https://doi.org/10.1016/j.molcel.2021.11.035
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/
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.
Recommended or similar items
The current recommendation prototype on the Apollo Repository will be turned off on 03 February 2023. Although the pilot has been fruitful for both parties, the service provider IKVA is focusing on horizon scanning products and so the recommender service can no longer be supported. We recognise the importance of recommender services in supporting research discovery and are evaluating offerings from other service providers. If you would like to offer feedback on this decision please contact us on: support@repository.cam.ac.uk