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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

JoVE Journal
Genética

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JoVE Journal Genética

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

•

11:35 min

•

June 16, 2017

•

Erin L. Sternburg, Kristen C. Dias, Fedor V. Karginov

¹Cell Biology and Neuroscience,University of California, Riverside

Capítulos

00:05Título
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

Tadução automática

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

Keywords: CRISPR/Cas9 Gene Knockout Nonhomologous End Joining Homology-directed Repair T-REx-293 Cells Transfection Drug Selection Clonal Isolation

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