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选择依赖和独立生成CRISPR / Cas9介导的哺乳动物细胞基因敲除
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Genetica
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JoVE Journal Genetica
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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选择依赖和独立生成CRISPR / Cas9介导的哺乳动物细胞基因敲除

DOI:
10.3791/55903-v

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11:35 min

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June 16, 2017

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, ,
1Cell Biology and Neuroscience,University of California, Riverside

Capitoli

  • 00:05Titolo
  • 00:57Transfection of CRISPER Component into Cultured Cells and Drug Selection
  • 02:31Isolation of Clonal Populations
  • 03:53Screening Candidates by Dot Blot
  • 06:30Screening Candidates by Colony PCR
  • 08:59Results: Knockout Cell Lines Generated by Two CRISPER/Cas9 Methods
  • 10:40Conclusion

Summary

Traduzione automatica

Traduzione automatica

遗传操纵体细胞系的能力的最新进展对于基础和应用研究具有巨大的潜力。在这里，我们提出了CRISPR / Cas9在哺乳动物细胞系中产生敲除生产和筛选的两种方法，使用和不使用选择标记。

Tags

CRISPR/Cas9Gene KnockoutNonhomologous End JoiningHomology-directed RepairT-REx-293 CellsTransfectionDrug SelectionClonal Isolation

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