TAT peptides mediated small interfering RNA delivery to Huh-7 cells and efficiently inhibited hepatitis C virus RNA replication.
OBJECTIVE
To observe whether or not the small-interfering RNA (siRNA) that conjugated with human immunodeficiency virus type 1 (HIV-1) TAT(47-57) peptides can enter Huh-7 cells and efficiently silence hepatitis C virus (HCV) infection in cell culture.


METHODS
siRNA targeting the highly conserved stem loop IV of the HCV 5' untranslated region (5'UTR) was conjugated to TAT(47-57) peptides via the crosslinker sulfosuccinimidyl-4-(p-maleimidophenyl)-butyrate, and then the conjugates were added to the Huh-7 cell culture. Firefly luciferase activity and HCV RNA were assessed using a luciferase assay reagent and real-time reverse transcript polymerase chain reaction, respectively.


RESULTS
The expression of firefly luciferase in HCV replicons and the concentration of HCV RNA were downregulated by siRNA-TAT(47-57), and siRNA-TAT(47-57) mediated RNA interfering activity which was directly correlated with increasing concentrations of the siRNA-TAT(47-57) conjugate used.


CONCLUSIONS
Cell-penetrating peptides such as HIV-1 TAT are an effective method for the delivery of siRNA targeted at 5'UTR of HCV in mammalian cells.
Meng
Shuang
S
National Center for Clinical Laboratories, Beijing Hospital, Beijing, PR China.
Wei
Baojun
B
Xu
Ruihuan
R
Zhang
Kuo
K
Wang
Lunan
L
Zhang
Rui
R
Li
Jinming
J
D020121:5' Untranslated Regions; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D020868:Gene Silencing; D017930:Genes, Reporter; D016174:Hepacivirus / Q000187:drug effects*; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014779:Virus Replication / Q000187:drug effects*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000494:pharmacology*
D020121:5' Untranslated Regions; D000998:Antiviral Agents; D034741:RNA, Small Interfering; D054322:tat Gene Products, Human Immunodeficiency Virus; D049409:Luciferases, Firefly