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Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P-associated

Qiudi Deng1, Yujun Liu2,3, Xin Li1

  • 1Department of Biotechnology, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong 510632, China.

RNA (New York, N.Y.)
|February 27, 2019
PubMed

Insights

Engineered external guide sequences (EGSs) efficiently target and degrade human cytomegalovirus mRNA, significantly reducing viral growth. This EGS variant shows promise as a novel gene-targeting therapy for HCMV infections.

Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • External guide sequences (EGSs) are short RNAs that direct ribonuclease P (RNase P) to cleave specific mRNAs.
  • EGS technology offers potential for gene-targeting therapies.
  • Previous studies demonstrated engineered EGS variants can effectively hydrolyze target mRNAs.

Purpose of the Study:

  • To design and evaluate an EGS variant targeting the mRNA of human cytomegalovirus (HCMV) major capsid protein (MCP).
  • To assess the efficiency of the engineered EGS variant in inhibiting HCMV gene expression and viral replication.

Main Methods:

  • An EGS variant complementary to HCMV MCP mRNA was designed.
  • In vitro assays measured RNase P-mediated mRNA cleavage efficiency.
  • Cell-based experiments assessed MCP expression and viral growth reduction.

Main Results:

  • The engineered EGS variant was 80-fold more efficient than natural tRNA-derived EGS in cleaving HCMV MCP mRNA in vitro.
  • In cells, the EGS variant reduced MCP expression by 98% and viral growth by 10,000-fold.
  • Natural tRNA-originated EGS reduced MCP expression by 73-74% and viral growth by 200-fold.

Conclusions:

  • Engineered EGS variants demonstrate superior efficiency in inhibiting HCMV gene expression and viral growth compared to natural tRNA-derived EGS.
  • The developed EGS variant is a potent candidate for treating HCMV infections.
Keywords:
RNase Pexternal guide sequencegene-targeted therapyhuman cytomegalovirus

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