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Chemical Dimerization-Induced Protein Condensates on Telomeres
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Biologia
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JoVE Journal Biologia
Chemical Dimerization-Induced Protein Condensates on Telomeres

Chemical Dimerization-Induced Protein Condensates on Telomeres

DOI:
10.3791/62173-v

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08:52 min

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April 12, 2021

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, ,
1Department of Biological Sciences, Mellon College of Science,Carnegie Mellon University, 2Department of Chemistry, School of Arts and Sciences,University of Pennsylvania

Capitoli

  • 00:04Introduction
  • 00:37Dimerization on Telomeres
  • 01:11Immunofluorescence (IF)
  • 03:02Fluorescence In Situ Hybridization (FISH)
  • 04:14Imaging
  • 06:23Results: Chemical Dimerization to Induce Chromatin-associated Condensates
  • 08:06Conclusion

Summary

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This protocol illustrates a chemically induced protein dimerization system to create condensates on chromatin.  The formation of promyelocytic leukemia (PML) nuclear body on telomeres with chemical dimerizers is demonstrated. Droplet growth, dissolution, localization and composition are monitored with live cell imaging, immunofluorescence (IF) and fluorescence in situ hybridization (FISH).

Tags

Chemical DimerizationProtein CondensatesTelomeresChromatinLive Cell ImagingBiochemical AssaysHalo-GFP-TRF1MCherry-eDHFR-SIMMCherry-eDFR-SIMPMLSUMO-1SUMO-2SUMO-3DAPIImmunofluorescence

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