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DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation
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Biologia
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JoVE Journal Biologia
DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation

DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation

DOI:
10.3791/1520-v

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15:56 min

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October 19, 2009

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, , ,
1Department of Physiology,David Geffen School of Medicine, University of California, Los Angeles

Capitoli

  • 00:00Titolo
  • 01:16Introduction
  • 01:49Plasmid Preparation for Electroporation
  • 02:46Animal Preparation for Electroporation
  • 06:12Electroporation and Transfection
  • 08:31Assaying In-Vivo Transfection by Electroporation
  • 08:36Transfection Efficiency
  • 09:47Subcellular Protein Localization
  • 12:31Expression and Targeting of the a-sub-unit of the Dihydropyridine Receptor (DHPR-a1s)
  • 13:56Functional Evaluation of the Expression of EYFP-CIC1
  • 15:30Conclusion

Summary

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We describe detailed procedures for the efficient transfection of plasmid DNA into the fibers of foot muscles of live mice using electroporation and the subsequent visualization of protein expression using fluorescence microscopy.

Tags

DNA TransfectionMammalian Skeletal MusclesIn Vivo ElectroporationCell BiologyTransgenic ProteinsFluorescently Tagged ConstructsMutant VariantsIn Vivo TransfectionPlasmidsLiverSkeletal MuscleBrainFlexor Digitorum BrevisInterosseus MusclesAdult MiceExperimental ParametersSoluble ProteinsStructural ProteinsMembrane ProteinsEGFPECFP3CalpainFKBP12β2a-DHPRMinidystrophinα-actininα1s-DHPRRyR1Cardiac Na/Ca2+ ExchangerNaV1.4 Na ChannelSERCA1

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