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Værktøjer til at undersøge, hvilken rolle for Arkitektonisk Protein HMGB1 i behandling af Helix fordrejende, site-specifikke DNA interstrengstværbindinger
Journal JoVE
Génétique
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Journal JoVE Génétique
Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks
DOI:

12:19 min

November 10, 2016

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Chapitres

  • 00:05Titre
  • 00:50Transfection of TFO-directed ICL-containing Plasmids in Human Cells
  • 02:27Crosslinking and DNA Isolation
  • 05:21Chromatin Immunoprecipitation of TFO-directed ICL-containing Plasmids
  • 07:06DNA Supercoiling Assay and Two Dimensional Agarose Gel Electrophoresis
  • 09:30Studying DNA Topology Modification Using ChIP and DNA Supercoiling Assays
  • 10:45Conclusion

Summary

Traduction automatique

Targeted DNA damage can be achieved by tethering a DNA damaging agent to a triplex-forming oligonucleotide (TFO). Using modified TFOs, DNA damage-specific protein association, and DNA topology modification can be studied in human cells by the utilization of modified chromatin immunoprecipitation assays and DNA supercoiling assays described herein.

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