RNase P-mediated inhibition of viral growth by exogenous administration of short oligonucleotide external guide sequence.
The use of external guide sequence (EGS) in directing endogenous ribonuclease P (RNase P) for inhibition of viral propagation is described in this chapter, with an emphasis on chemically modified EGSs and their extracellular delivery. Targeting of the mRNA-encoding human cytomegalovirus (HCMV) protease by DNA-based EGSs is presented as an example of how to design chemically modified EGSs for antiviral applications. General information about the EGS-based technology is included, followed by detailed protocols for EGS design, human RNase P purification, in vitro assay of EGS activity, liposome-mediated delivery of chemically modified EGSs and detection of their distribution in cells, and an assay of EGS activity for blocking growth of HCMV in cultured cells.
Dunn
Walter
W
Division of Infectious Diseases, School of Public Health, University of California, Berkeley, USA.
Liu
Fenyong
F
D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002852:Chromatography, Ion Exchange; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D012333:RNA, Messenger / Q000737:chemistry; D012365:RNA, Transfer, Tyr / Q000235:genetics; D043262:Ribonuclease P / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D014780:Viruses / Q000187:drug effects / Q000254:growth & development*
D000998:Antiviral Agents; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012365:RNA, Transfer, Tyr; D043262:Ribonuclease P