Characterization of Escherichia coli RNase H Discrimination of DNA Phosphorothioate Stereoisomers.

ABSTRACT: Phosphorothioate (PS) modification of antisense oligonucleotides (ASOs) is a critical factor enabling their therapeutic use. Standard chemical synthesis incorporates this group in a stereorandom manner; however, significant effort was made over the years to establish and characterize the impact of chiral control. In this work, we present our in-depth characterization of interactions between Escherichia coli RNase H and RNA-DNA heteroduplexes carrying chirally defined PS groups. First, using a massive parallel assay, we showed that at least a single Rp-PS group is necessary for efficient RNase H-mediated cleavage. We followed by demonstrating that this group needs to be aligned to the phosphate-binding pocket of RNase H, and that chiral status of other PS groups in close proxi

SUBMITTER: Kielpinski LJ 

PROVIDER: S-EPMC8713576 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature