posted on 2024-02-27, 19:14authored byLiang-Liang Wang, Chao-Qun Wu, Qiu-Long Zhang, Yang Wang, Yan Liu, Wen-Jian Yang, Sen-Lin Ye, Yongqiang Tian, Liang Xu
RNA-cleaving ribozymes are promising candidates as general
tools
of RNA interference (RNAi) in gene manipulation. However, compared
with other RNA systems, such as siRNA and CRISPR technologies, the
ribozyme tools are still far from broad applications on RNAi due to
their poor performance in the cellular context. In this work, we report
an efficient RNAi tool based on chemically modified hammerhead ribozyme
(HHR). By the introduction of an intramolecular linkage into the minimal
HHR to reconstruct the distal interaction within the tertiary ribozyme
structure, this cross-linked HHR exhibits efficient RNA substrate
cleavage activities with almost no sequence constraint. Cellular experiments
suggest that both exogenous and endogenous RNA expression can be dramatically
knocked down by this HHR tool with levels comparable to those of siRNA.
Unlike the widely applied protein-recruiting RNA systems (siRNA and
CRISPR), this ribozyme tool functions solely on RNA itself with great
simplicity, which may provide a new approach for gene manipulation
in both fundamental and translational studies.