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

Mathilde Beaufils; Amandine Tourel; Anne Petiot; Nicole B. Halmai; Dave J. Segal; John Rendu; Isabelle Marty

doi:10.3791/64114

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Génétique

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

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Journal JoVE Génétique

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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Mathilde Beaufils, Amandine Tourel, Anne Petiot, Nicole B. Halmai, Dave J. Segal, John Rendu, Isabelle Marty

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

Chapitres

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

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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.

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

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Chapitres

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