Engineering an adenine base editor in human embryonic stem cells with minimal DNA and RNA off-target activities.

ABSTRACT: Genome editing in pluripotent stem cells (PSCs) using CRISPR technology holds great promise for therapeutic applications. Yet, it has been reported that Cas9-mediated cleavage could cause large deletions or rearrangements of DNA, and the selection of edited PSCs could acquire p53 mutations. Adenine base editors (ABEs) do not introduce DNA double-strand breaks and thus have been proposed as alternatives to circumvent those problems, but their off-target effects still limit their applications. Here, we tested different combinations of off-target reduction methods to further diminish off-target effects of ABEs without compromising their on-target editing efficiencies. We subsequently chose the best editor, CE-8e-dV, which contains V106W substitution, R153 deletion, and Cas-embedding strategy,

SUBMITTER: Zhang Z 

PROVIDER: S-EPMC9375152 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature