A/C Simultaneous Conversion Using the Dual Base Editor in Human Cells

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

Base editors, mostly cytidine base editors (CBEs) and adenine base editors (ABEs), are powerful tools for precise base editing. However, current base editors can only edit either adenines or cytosines. Thus, our lab has developed a dual base editor (A&C-BEmax) through the fusion of cytidine and adenosine deaminases to Cas9n to achieve both C•G to T•A and A•T to G•C mutations, which enables A/C simultaneous conversion in the same allele (up to 30%) in human cells. Here, we described a protocol for the usage of A&C-BEmax in human cells. This protocol includes standard dual base editing experiments in HEK293T cells and data analysis of dual base editing outcomes using BE-analyzer. All the workflow of experiments can be completed within 2–3 weeks.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages63-72
Number of pages10
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2606
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Adenine deaminase
  • Base editing
  • Cas9
  • Cytosine

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