Engineering of highly active gene targeting RNase P ribozyme against human cytomegalovirus infection
Year: 2025
Type: article
Abstract: Sequence-specific ribonuclease (RNase) P ribozymes can be engineered in vitro and are promising gene-targeting agents to knock down gene expression. In this study, we applied an RNase P ribozyme variant to hydrolyze the mRNA of human cytomegalovirus (HCMV) major capsid protein (MCP), which is necessary for viral capsid formation and growth. Functional variant R668-F was about 100 times more efficient in slicing the MCP mRNA in vitro than M1-F, the ribozyme with a natural RNase P ribozyme sequence. In R668-F-expressing cells, a decrease of about 98%–99% in the expression of MCP was detected, and the virus production was reduced by 70,000 folds. However, the expression of inactive control ribozymes in cells resulted in a less than 10% decrease in MCP expression and no apparent decrease in HCMV growth. In cells observed with the ribozyme-mediated reduction of MCP expression , HCMV capsid formation and virus growth were inhibited and the expressions of other viral genes were unaffected. These findings provide the first direct evidence that ribozyme R668-F specifically inhibits MCP expression and blocks HCMV growth. Our results further suggest that the engineered RNase P ribozymes , including R668-F, may act as a novel general gene-targeting strategy to treat infections of viruses, including HCMV.
Source: hLife
Authors: Yujun Liu, Bin Yan, Isadora Zhang, Fenyong Liu
Institution: University of California, Berkeley
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,300
OpenAlex ID: W4407953684