Hepatitis C virus replication-specific inhibition of microRNA activity with self-cleavable allosteric ribozyme.
Functional sequestration of microRNA 122 (miR-122) by treatment with an oligonucleotide complementary to the miRNA results in long-lasting suppression of hepatitis C virus (HCV) viremia in primates. However, the safety of the constitutive miR-122 silencing approach to HCV inhibition is unclear, since miR-122 can modulate the expression of many host genes. In this study, a regulation system capable of specifically inhibiting miR-122 activity only upon HCV infection was developed. To this end, an allosteric self-cleavable ribozyme capable of releasing antisense sequence to miR-122 only in the presence of HCV nonstructural protein 5B was developed using in vitro selection method. The activity of the reporter construct with miR-122 target sequences at its 3' untranslated region and the expression of endogenous miR-122 target proteins were specifically stimulated through sequestration of miR-122 only in HCV replicon Huh-7 cells, but not in naïve Huh-7 cells, when transfected with expression vector encoding the specific allosteric ribozyme. These findings indicate that miR-122 function can be specifically inhibited by the allosteric ribozyme only in HCV-replicating cells. Importantly, HCV replicon replication was efficiently inhibited by the allosteric ribozyme. This ribozyme could be useful for the specific, safe, and efficacious anti-HCV modulation.
Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Yongin, Republic of Korea.
Chang Ho
CH
Lee
Ji Hyun
JH
Kim
Hee Won
HW
Kim
Heejoon
H
Myung
Seong-Wook
SW
Lee
D000494:Allosteric Regulation; D001483:Base Sequence; D017931:DNA Primers; D016174:Hepacivirus / Q000502:physiology*; D035683:MicroRNAs / Q000037:antagonists & inhibitors*; D016337:RNA, Catalytic / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D014779:Virus Replication
D017931:DNA Primers; D035683:MicroRNAs; D016337:RNA, Catalytic