Enhanced hammerhead ribozyme turnover rates: Reevaluating therapeutic space for small catalytic RNAs.

ABSTRACT: We discovered enhanced hammerhead ribozyme (EhhRz) kinetic performance during therapeutic optimization of a lead agent against the human rhodopsin mRNA target. Turnover activity in trans was greater than 300 nMmin^-1 under 10-fold substrate excess and physiological Mg²⁺ (1 mM). We validated a moderate-throughput fluorescence quantitative hhRz kinetic assay, which is linear with substrate and product moles. The EhhRz targets a CUC↓ cleavage site in a substrate with no predicted secondary/tertiary structure and demonstrates classical Michaelis-Menten turnover behavior with efficiency (V _max/K _m ) up to 3.2 × 10⁶ min^-1M^-1. EhhRzs show cooperative Mg²⁺ titration (K

SUBMITTER: Myers JM 

PROVIDER: S-EPMC11875169 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature