A Cas9-mediated adenosine transient reporter enables enrichment of ABE-targeted cells.

ABSTRACT:

Background

Adenine base editors (ABE) enable single nucleotide modifications without the need for double-stranded DNA breaks (DSBs) induced by conventional CRIPSR/Cas9-based approaches. However, most approaches that employ ABEs require inefficient downstream technologies to identify desired targeted mutations within large populations of manipulated cells. In this study, we developed a fluorescence-based method, named "Cas9-mediated adenosine transient reporter for editing enrichment" (CasMAs-TREE; herein abbreviated XMAS-TREE), to facilitate the real-time identification of base-edited cell populations.

Results

To establish a fluorescent-based assay able to detect ABE activity within a cell in real time, we designed a construct encoding a mCherry fluorescent protein followed

SUBMITTER: Brookhouser N 

PROVIDER: S-EPMC7737295 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature