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Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation
JoVE Journal
Neurociência
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JoVE Journal Neurociência
Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

DOI:
10.3791/2821-v

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07:38 min

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June 28, 2011

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, ,
1Department of Pathology and Immunology,Washington University School of Medicine

Capítulos

  • 00:05Título
  • 01:30Eye Collection
  • 02:43Retinal Dissection
  • 03:58Preparation for Electroporation
  • 04:34Electroporation
  • 05:27Culturing of Electroporated Retinas
  • 06:23Visualization of cis-regulated Element (CRE) Expression in Electroporation Retinal Cells
  • 07:17Conclusion

Summary

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This protocol describes a simple and inexpensive way to quantify the activity of cis-regulatory elements (i.e., enhancer/promoters) in living mouse retinas via explant electroporation. DNA preparation, retinal dissection, electroporation, retinal explant culture, and post-fixation analysis and quantification are described.

Tags

Cis-regulatory ElementsMouse RetinaExplant ElectroporationTranscription FactorsGene NetworksEnhancersPromotersTranscriptional ActivatorsTranscriptional RepressorsSpatiotemporal PatternGene ExpressionGenomic DNATransgenesisEnhancer FunctionQuantitative AssaysIn Vivo RelevanceElectroporation

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