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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
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Cancer Research
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JoVE Journal Cancer Research
Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

DOI:
10.3791/61557-v

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09:22 min

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February 28, 2021

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, , , ,
1Department of Biomedical Sciences, Asan Medical Institute of Convergence Science and Technology, Asan Medical Center,University of Ulsan College of Medicine, 2Stem Cell Immunomodulation Research Center,University of Ulsan College of Medicine

Chapters

  • 00:05Introduction
  • 01:03Design and Construction of BRCA1 Targeting gRNAs
  • 03:17Creation of BRCA1 Variants using CRISPR-mediated Base Editing Tools
  • 04:15Sample Preparation for Illumina Next-Generation Sequencing (NGS)
  • 07:01Results: Functional Study of BRCA1 using CRISPR-Cas9 Systems
  • 08:34Conclusion

Summary

Automatic Translation

Automatic Translation

People with BRCA1 mutations have a higher risk of developing cancer, which warrants accurate evaluation of the function of BRCA1 variants. Herein, we described a protocol for functional assessment of BRCA1 variants using CRISPR-mediated cytosine base editors that enable targeted C:G to T:A conversion in living cells.

Tags

BRCA1CRISPRBase EditorsFunctional AssessmentPoint MutationsCancerDrug TargetsGenome SequenceGuide RNAOligonucleotidesPlasmid DNATransformationSanger SequencingHAP1-BE3 CellsTransfection

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