Abstract
Adenine base editors (ABEs) have been subjected to multiple rounds of mutagenesis with the goal of optimizing their function as efficient and precise genome editing agents. Despite this ever-increasing data accumulation of the effects that these mutations have on the activity of ABEs, the molecular mechanisms defining these changes in activity remain to be elucidated. In this study, we provide a systematic interpretation of the nature of these mutations using an entropy-based classification model that relies on evolutionary data from extant protein sequences. Using this model in conjunction with experimental analyses, we identify two previously reported mutations that form an epistatic pair in the RNA-editing functional landscape of ABEs. Molecular dynamics simulations reveal the atomistic details of how these two mutations affect substrate-binding and catalytic activity, via both individual and cooperative effects, hence providing insights into the mechanisms through which these two mutations are epistatically coupled.
Competing Interest Statement
A.C.K. is a member of the SAB and a consultant of Pairwise Plants, and is an equity holder for Pairwise Plants and Beam Therapeutics. A.C.K.'s interests have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. All other authors declare that they have no competing interests.