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Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays
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Cancer Research
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JoVE Journal Cancer Research
Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays

Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays

DOI:
10.3791/58710-v

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11:05 min

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January 07, 2019

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, , , , , , ,
1Tumor Initiation and Maintenance Program,Sanford Burnham Prebys Medical Discovery Institute, 2Blood Research Institute,Blood Center of Wisconsin, 3Department of Bioengineering,University of California, San Diego

Chapters

  • 00:04Title
  • 00:30Medium-throughput Cloning of sgRNAs
  • 04:55Viral Production of sgRNA Constructs and Transduction in AML-Cas9 Cells
  • 07:10Competitive Growth Assay
  • 09:21Results: The Role of Candidate Genes in AML Cell Proliferation
  • 10:30Conclusion

Summary

Automatic Translation

Automatic Translation

This manuscript describes a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) CRISPR-Cas9-based method for simple and expeditious investigation of the role of multiple candidate genes in Acute Myeloid Leukemia (AML) cell proliferation in parallel. This technique is scalable and can be applied in other cancer cell lines as well.

Tags

Keywords: CRISPR-Cas9Competition AssaysGenetic DependenciesAMLSingle Guide RNAOligonucleotidesAnnealingLigationVector LinearizationHigh-throughput Screening

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