Related Experiment Video
Updated: Oct 8, 2025
Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
Published on: May 30, 2012
TRACE generates fluorescent human reporter cell lines to characterize epigenetic pathways.
Iva A Tchasovnikarova1, Sharon K Marr2, Manashree Damle2
1Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Genetics, Harvard Medical School, Boston, MA 02114, USA; The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
We developed TRACE, a high-throughput method to create reporter cell lines for studying epigenetic pathways. This technique identifies genetic reporters responsive to epigenetic factor manipulation, advancing chromatin biology research.
Area of Science:
- Cell Biology
- Epigenetics
- Genomics
Background:
- Genetically encoded biosensors are crucial for monitoring cellular behavior.
- Generating reporters for chromatin factors is challenging, hindering epigenetic pathway dissection.
- Understanding epigenetic mechanisms requires effective tools to study chromatin dynamics.
Purpose of the Study:
- To present TRACE (transgene reporters across chromatin environments), a high-throughput, genome-wide technique.
- To generate fluorescent human reporter cell lines responsive to epigenetic factor manipulation.
- To enable genetic interrogation of epigenetic pathways and advance chromatin biology.
Main Methods:
- Profiling GFP expression from barcoded lentiviral integrants under varying conditions.
- Utilizing a large pool of individually barcoded lentiviral integrants.
- Manipulating HIV-1 host factor LEDGF to alter lentiviral integration site preferences.
Main Results:
- Identified reporters responsive to LSD1 inhibition and SUZ12 ablation.
- Demonstrated TRACE's ability to generate cell lines sensitive to epigenetic perturbations.
- Expanded the range of chromatin environments examined by TRACE through LEDGF manipulation.
Conclusions:
- TRACE provides a powerful platform for generating phenotypic reporters for epigenetic studies.
- This technique facilitates the genetic interrogation of diverse epigenetic pathways.
- TRACE will significantly advance mechanistic understanding of chromatin biology.
