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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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JoVE Journal Genetik
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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

Kapitel

  • 00:05Titel
  • 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

Automatische Übersetzung

Automatische Übersetzung

Recent advances in the ability to genetically manipulate somatic cell lines hold great potential for basic and applied research. Here, we present two approaches for CRISPR/Cas9 generated knockout production and screening in mammalian cell lines, with and without the use of selectable markers.

Tags

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

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