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Targeting mRNAs by engineered sequence-specific RNase P ribozymes

Yong Bai1, Naresh Sunkara, Fenyong Liu

  • 1Division of Infectious Diseases and Vaccinology, School of Public Health, University of California, Berkeley, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 9, 2012
PubMed

Insights

Engineered RNase P catalytic RNA (M1GS RNA) effectively targets and cleaves human cytomegalovirus (HCMV) mRNA in cells. This ribozyme therapy significantly reduces viral gene expression and inhibits viral growth.

Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Engineered ribozymes offer a novel approach for gene-specific targeting.
  • RNase P catalytic RNA (M1GS RNA) can be customized for therapeutic applications.

Purpose of the Study:

  • To describe methods for using M1GS RNA for in vitro and in vivo trans-cleavage of viral mRNA.
  • To evaluate the antiviral activity of a customized M1GS ribozyme against human cytomegalovirus (HCMV).

Main Methods:

  • Mapping accessible viral mRNA regions in infected cells.
  • Generating and assaying customized M1GS ribozymes for in vitro cleavage.
  • Assessing M1GS RNA expression and antiviral efficacy in cultured cells.

Main Results:

  • A functional M1GS ribozyme was constructed to cleave HCMV mRNAs encoding capsid scaffolding protein (CSP) and assemblin.
  • In HCMV-infected human cells, M1GS RNA reduced CSP and assemblin expression by over 85%.
  • A 4,000-fold reduction in viral growth was observed in cells expressing the M1GS ribozyme.

Conclusions:

  • Customized M1GS ribozymes serve as effective gene-targeting agents.
  • M1GS ribozymes demonstrate potent in trans-cleavage activity against viral genes.
  • This approach shows promise for inhibiting viral growth and developing antiviral therapies.

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