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Base Editors pp 73–85Cite as

Functional Analysis of Variants in BRCA1 Using CRISPR Base Editors

Part of the Methods in Molecular Biology book series (MIMB,volume 2606)

Abstract

To date, methods such as fluorescent reporter assays, embryonic stem cell viability assays, and therapeutic drug-based sensitivity assays have been used to evaluate the function of the variants of uncertain significance (VUS) of the BRCA genes. However, these methods have limitations as they are associated with overexpression and do not apply to post-transcriptional regulation. Therefore, there are several VUS whose functions are unclear. Recently, we devised a new way to assess the functionality of variants in BRCA1 via a CRISPR-mediated base editor to overcome these limitations. We precisely introduced the target nucleotide substitution in living cells and identified variants whose functions were not defined. Here, we describe the methods for the functional appraisal of BRCA1 variants using CRISPR-based base editors.

Key words

  • CRISPR
  • Base editing
  • Genome engineering
  • Functional assessment
  • BRCA1
  • Cytosine base editor
  • Adenine base editor

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Acknowledgments

This study was supported by the National Research Foundation of Korea (grants 2017M3A9B4062419, 2020R1F1A1075508, 2018R1A5A2020732, and 2021R1C1C1007162 to YK and 2021R1A6A3A13046696 to J.-E.S.).

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Authors and Affiliations

  1. Department of Biomedical Sciences, Asan Medical Institute of Convergence Science and Technology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea

    Ji-Eun See & Yongsub Kim

  2. Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea

    Ji-Eun See & Yongsub Kim

Authors
  1. Ji-Eun See
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  2. Yongsub Kim
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Editor information

Editors and Affiliations

  1. Seoul National University College of Medicine, Seoul, Korea (Republic of)

    Sangsu Bae

  2. Seoul National University College of Medicine, Seoul, Korea (Republic of)

    Beomjong Song

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See, JE., Kim, Y. (2023). Functional Analysis of Variants in BRCA1 Using CRISPR Base Editors. In: Bae, S., Song, B. (eds) Base Editors. Methods in Molecular Biology, vol 2606. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2879-9_7

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  • DOI: https://doi.org/10.1007/978-1-0716-2879-9_7

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