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Development of Knock-Out Muscle Cell Lines using Lentivirus-Mediated CRISPR/Cas9 Gene Editing
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JoVE Journal Genetics
Development of Knock-Out Muscle Cell Lines using Lentivirus-Mediated CRISPR/Cas9 Gene Editing

Development of Knock-Out Muscle Cell Lines using Lentivirus-Mediated CRISPR/Cas9 Gene Editing

DOI:
10.3791/64114-v

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10:12 min

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June 16, 2022

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, , , , , ,
1University Grenoble Alpes, INSERM, U1216, CHU Grenoble Alpes, Grenoble Institut Neurosciences, 2Genome Center,University of California, Davis

Chapters

  • 00:05Introduction
  • 00:43CRISPR Guide Design
  • 02:03Plasmid Cloning
  • 03:43Lentivirus Production
  • 06:28Myoblast Transduction
  • 07:20Functional Characterization of Edited Clones
  • 08:53Results: Development of RYR1-Knock Out Human Muscle Cell Line
  • 09:42Conclusion

Summary

Automatic Translation

Automatic Translation

The protocol describes how to generate knock-out myoblasts using CRISPR/Cas9, starting from the design of guide-RNAs to the cellular cloning and characterization of the knock-out clones.

Tags

Keywords: CRISPR/Cas9Gene EditingKnock-outMuscle CellsLentivirusIPS CellsGuide RNAPCRAgarose GelRestriction EnzymesPlasmid Cloning

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