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Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions
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JoVE Journal 遺伝学
Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions

Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions

DOI:
10.3791/55919-v

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10:10 min

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May 28, 2017

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, , , , , , ,
1Center for Genome Research, Department of Life Sciences,University of Modena and Reggio Emilia, 2Laboratory of Chromatin and Gene Regulation During Development,Imagine Institute, 3Institute for Biomedical Technologies,CNR, 4Généthon, 5Sorbonne Paris Cité – Université Paris Descartes

章

  • 00:05標題
  • 00:45MLV Transduction of Human Cells
  • 01:41Restriction Enzyme Digestion and Linker Ligation
  • 03:05First-round Linker-mediated PCR
  • 03:43Second Round Linker-mediated PCR
  • 05:17Library Preparation and Dilution for Massive Sequencing of MLV Integration Sites
  • 07:53Results: Mapping MLV Integration into Epigenetically-defined Regulatory Regions
  • 09:23Conclusion

概要

自動翻訳

自動翻訳

Here, we describe a protocol for genome-wide mapping of the integration sites of Moloney murine leukemia virus-based retroviral vectors in human cells.

タグ

Retroviral ScanningMLV Integration SitesCell-specific Regulatory RegionsGene TherapyGene RegulationMLV-derived Retroviral VectorEGFP Reporter GeneFlow Cytometric AnalysisGenomic DNA PreparationRestriction Enzyme DigestionLinker LigationFirst-round PCRSecond-round PCR

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