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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
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Journal JoVE Biochimie
Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

DOI:
10.3791/62949-v

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06:07 min

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August 05, 2022

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, ,
1Department of Chemistry,Bucknell University, 2Program in Cell Biology/Biochemistry,Bucknell University

Chapitres

  • 00:05Introduction
  • 00:24Prepare Assay Solutions for the Screen
  • 01:48Prepare Enzyme Solutions for the Screen
  • 02:52Run the Methylation Assay and Analyze the Data
  • 04:29Results: Determination of Initial Velocities of DNA Methylation in the Presence and Absence of Potential Inhibitors
  • 05:40Conclusion

Summary

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Traduction automatique

DNA methyltransferases are potential cancer drug targets. Here, a protocol is presented to assess small molecules for DNA methyltransferase inhibition. This assay utilizes an endonuclease to couple DNA methylation to fluorescence generation and allows for enzyme activity to be monitored in real time. 

Tags

Keywords: Continuous Fluorescence-based AssayEndonuclease-coupledDNA Methyltransferase InhibitorsPlate Reader AssayReal-time Data CollectionDNA MethylationSAMGlaIDNMT1DNA SubstrateInhibitor Screening

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