Evaluation of DNA Minicircles for Delivery of Adenine Base Editors Using Activatable Reporter Imaging Systems
Abstract
Base editing is a powerful genome editing tool with the potential to treat thousands of genetic diseases caused by single-base changes in DNA called point mutations. Adenine base editors (ABEs) specifically convert adenine (A) to guanine (G) and can theoretically correct almost half of all disease-causing point mutations. Currently, there is a need for a safe and efficient method of delivering ABEs into tissues. In Chapter 2, we developed DNA minicircles as a novel non-viral delivery method for ABEs. To assess delivery efficiency, we also developed activatable reporter imaging systems for the visualization of ABE activity using fluorescence or bioluminescence imaging. Using our imaging reporters, we show for the first time that minicircles are superior to plasmids for the delivery of ABE into a variety of cancer cell types. Chapter 3 summarizes the main findings and discusses limitations and future work.