TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways.
Tchasovnikarova, Iva A
Marr, Sharon K
Kingston, Robert E
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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
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.
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
Damon Runyon Cancer Research Foundation NIH
Damon Runyon Cancer Research Foundation (42-20)
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
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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