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

AbstractBase 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.Key wordsBase editingCas9Adenine deaminaseCytosine