Nanobodies for detecting and manipulating A to I editing enzymes and their modified RNA products

DESCRIPTION (provided by applicant): Adenosine to inosine or A-to-I RNA editing executed by ADAR family proteins is abundant in the central nervous system (CNS). The availability of tools to monitor and manipulate both ADAR enzymes and A-to-I RNA modifications will facilitate investigation of the relationships between specific A-to-I conversions and their function in CNS development. The goal of this project is to develop a set of nanobodies to A-to-I RNA editing enzymes and their modified RNA products. These nanobodies are expected to serve not only as reagents for detection, quantification, or immunoprecipitation but also as intracellular antibodies capable of inhibiting the activity of target ADAR enzymes or binding to specific A-to-I modified RNAs in cells. As proof-of-principle, in Phase I, using Abzyme's proprietary self-diversifying single domain antibody library platform for high throughput antibody discovery, nanobodies to inosine-containing generic RNAs and site- specific A-to-I mRNA isoforms