Journal
/
Genetics
/
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
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

•

11:35 min

•

June 16, 2017

•

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/55903?article_type=v

Parallel Measurement of Circadian Clock Gene Expression and Hormone Secretion in Human Primary Cell Cultures
check_url/55903?article_type=v

Rearing and Double-stranded RNA-mediated Gene Knockdown in the Hide Beetle, Dermestes maculatus
check_url/55903?article_type=v

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
check_url/55903?article_type=v

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
check_url/55903?article_type=v

Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning
check_url/55903?article_type=v

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
check_url/55903?article_type=v

A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
check_url/55903?article_type=v

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
check_url/55903?article_type=v

Formaldehyde-assisted Isolation of Regulatory Elements to Measure Chromatin Accessibility in Mammalian Cells
check_url/55903?article_type=v

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

Read Article