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Direkter Gene Knock-out von Axolotl Spinal Cord Neural Stem Cells via Electroporation of CAS9 Protein-gRNA Complexes
JoVE Journal
Genetics
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JoVE Journal Genetics
Direct Gene Knock-out of Axolotl Spinal Cord Neural Stem Cells via Electroporation of CAS9 Protein-gRNA Complexes
DOI:

09:10 min

July 09, 2019

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Chapters

  • 00:04Title
  • 01:16CAS9-gRNA Ribonucleoprotein Mixture and Agarose Plate Preparation
  • 02:33Electroporator Configuration and Glass Microinjection Capillary Loading
  • 04:09Pneumatic Pump Configuration
  • 04:37CAS9-gRNA Ribonucleoprotein Mixture Injection
  • 06:05Electroporation
  • 06:45Knock-Out Efficiency Assessment
  • 07:25Results: Representative Knock-Out Effects in Axolotl Spinal Cord Neural Stem Cells
  • 08:33Conclusion

Summary

Automatic Translation

Hier wird ein Protokoll zur Durchführung zeit- und raumbeschränkter Gen-Knock-out in Axolotl-Rückenmarksen vorgestellt, indem der CAS9-gRNA-Komplex in den Zentralkanal des Rückenmarks injiziert wird, gefolgt von Elektroporation.

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