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Ribozyme inhibition of alphavirus replication
Attila A Seyhan1, Danielle Vitiello, Michele T Shields
1Somagenics, Inc., Santa Cruz, California 95060, USA.
The Journal of Biological Chemistry
|May 15, 2002
Summary
Researchers developed a system to test trans-acting ribozymes for antiviral activity against Sindbis virus. Active ribozymes significantly inhibited viral replication in mammalian cells, suggesting their potential as antiviral agents.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Trans-acting ribozymes are catalytic RNA molecules with potential therapeutic applications.
- Developing effective delivery and expression systems for ribozymes in mammalian cells is crucial for their use as antiviral agents.
Purpose of the Study:
- To develop and evaluate a model system for assessing the expression and antiviral activity of trans-acting ribozymes in mammalian cells.
- To engineer hairpin ribozymes targeting Sindbis virus RNA for antiviral therapy.
Main Methods:
- Engineered hairpin ribozymes targeting Sindbis virus RNA using activity-based selection.
- Developed transient and stable mammalian cell expression systems for ribozymes.
- Assessed ribozyme activity via RNA cleavage assays and viral replication inhibition using plaque and GFP assays.
Main Results:
- Ribozymes were expressed in mammalian cells with detectable RNA cleavage activity.
- Cells expressing active antiviral ribozymes showed significant inhibition of Sindbis virus replication.
- Control cells expressing inactive ribozymes showed minimal inhibition, validating ribozyme specificity.
Conclusions:
- The developed model system effectively demonstrates ribozyme-mediated inhibition of viral replication.
- Trans-acting ribozymes show promise as novel antiviral agents against RNA viruses like Sindbis virus.