Suppressing Kaposi's Sarcoma-Associated Herpesvirus Lytic Gene Expression and Replication by RNase P Ribozyme

Area of Science:

• Molecular Biology
• Virology
• Biochemistry

Background:

• Kaposi's sarcoma-associated herpesvirus (KSHV) causes Kaposi's sarcoma, an AIDS-defining illness.
• KSHV is an oncogenic virus critical for viral gene expression and replication.

Purpose of the Study:

• To engineer ribonuclease P (RNase P) ribozymes targeting KSHV RTA mRNA.
• To evaluate the efficacy of F-RTA ribozymes in suppressing KSHV replication and gene expression.

Main Methods:

• Engineering of RNase P ribozymes targeting KSHV RTA mRNA.
• In vitro cleavage assays of RTA mRNA by F-RTA ribozymes.
• In cellulo assessment of KSHV production and RTA expression upon ribozyme delivery.

Main Results:

• The F-RTA ribozyme efficiently cleaved RTA mRNA in vitro.
• Expression of F-RTA significantly suppressed KSHV production (250-fold) and RTA expression (92-94%) in cells.
• Control ribozymes showed minimal impact on KSHV and RTA expression.

Conclusions:

• RNase P ribozymes targeting RTA mRNA offer a potent strategy for KSHV suppression.
• F-RTA ribozymes effectively inhibit KSHV early and late gene expression, viral growth, and production.
• This represents a potential novel therapeutic approach for KSHV-associated diseases.