Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 175, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/62974
Keywords
-
Categories
Funding
- NSFC [31670762]
Ask authors/readers for more resources
The article discusses the importance of fusion genes in tumors, as well as the mechanism of action of the EWS-FLI1 fusion gene in Ewing sarcoma, including the formation of biomolecular condensates to enhance gene transcription. The DNA Curtains method can be used to visualize the assembly process of EWS-FLI1 condensates.
The fusion genes resulting from chromosomal translocation have been found in many solid tumors or leukemia. EWS-FLI1, which belongs to the FUS/EWS/TAF15 (FET) family of fusion oncoproteins, is one of the most frequently involved fusion genes in Ewing sarcoma. These FET family fusion proteins typically harbor a lowcomplexity domain (LCD) of FET protein at their N-terminus and a DNA-binding domain (DBD) at their C-terminus. EWS-FLI1 has been confirmed to form biomolecular condensates at its target binding loci due to LCD-LCD and LCD-DBD interactions, and these condensates can recruit RNA polymerase II to enhance gene transcription. However, how these condensates are assembled at their binding sites remains unclear. Recently, a single-molecule biophysics method-DNA Curtains-was applied to visualize these assembling processes of EWS-FLI1 condensates. Here, the detailed experimental protocol and data analysis approaches are discussed for the application of DNA Curtains in studying the biomolecular condensates assembling on target DNA.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available