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设计，包装，并通过立体定向注射高效价CRISPR复古和慢交付
JoVE Journal
Neurociência
This content is Free Access.
JoVE Journal Neurociência
Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection
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设计，包装，并通过立体定向注射高效价CRISPR复古和慢交付

DOI:
10.3791/53783-v

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11:28 min

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May 23, 2016

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, , , ,
1Department of Physiology and Neurobiology,Geisel School of Medicine at Dartmouth College

Capítulos

  • 00:05Título
  • 00:45Prepare 293FT/293GP Cells for Transfection
  • 03:02Calcium Phosphate Transfection and Viral Particle Collection
  • 05:24Concentration and Purification of the Virus
  • 06:42Stereotaxic Injection
  • 09:40Resultados
  • 10:59Conclusion

Summary

Tadução automática

Tadução automática

该CRISPR / Cas9系统提供作出有针对性的基因组编辑获得和负担得起科学界的潜力。该协议旨在演示如何创建病毒将基因敲除使用CRISPR / Cas9系统感兴趣的基因，然后立体定位将它们注入成年小鼠大脑。

Tags

CRISPRRetrovirusLentivirusStereotaxic InjectionNeurociênciaGene ManipulationGuide RNAPlasmid DesignCell CultureTransfectionHEK293T Cells

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