Leveraging Protein And gRNA Engineering Of Diverse CRISPR Systems To Optimize Adenosine Base Editor Activity For Correction Of A Causative Monogenic Disease Mutation

By Sean Crosson, Senior Scientist, Translational Biology

Optimizing adenine base editors for therapeutic single-nucleotide polymorphism (SNP) correction requires addressing both editing efficiency and off-target activity. Screening diverse CRISPR systems across disease-relevant loci identifies optimal lead architectures with favorable protospacer adjacent motif (PAM) requirements. Systematic, parallel engineering of protein components—including the PAM-interacting domain, nuclear localization signals, and base editing architectures—substantially increases targeted adenosine-to-guanosine conversion rates. Concurrently, refining the guide RNA backbone through sequence shortening and chemical modifications enhances potency while lowering required RNA doses. Introducing specialized deaminase variants further minimizes unwanted bystander activity, unintended cytosine editing, and off-target genomic cleavage. Combining these structural and enzymatic modifications converts baseline, low-efficiency editing frameworks into highly potent, specific tools capable of robust target correction exceeding 85%.

Explore the research data to learn how dual protein and guide engineering optimizes base editors for targeted genetic therapies.