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Effects of short RNA structural analogues against hepatitis C virus genotypes 2, 3 and 4 in replicon cells
Author(s) -
Elshaffei Ismail M,
Gupta Nidhi,
Wu Catherine H,
Wu David C,
Hammad Lamiaa N,
AboElmatty Dina M,
Mesbah Noha M,
Wu George Y
Publication year - 2015
Publication title -
journal of digestive diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 51
eISSN - 1751-2980
pISSN - 1751-2972
DOI - 10.1111/1751-2980.12250
Subject(s) - replicon , untranslated region , genotype , virology , transfection , luciferase , microbiology and biotechnology , biology , subgenomic mrna , rna , hepatitis c virus , stem loop , plasmid , viral replication , gene , virus , genetics
Objective To determine whether computer‐predicted short RNA structural analogues could inhibit hepatitis C virus ( HCV ) genotype 2a, 3a and 4a replication in cultured cells. Methods Short RNA sequences, X 12, X 12a and X 12b, designed to be identical in secondary structure to the X region in the 3′‐untranslated region (3′‐ UTR ) of the HCV 1b genome, as well as shorter stem‐loop components of X region, were inserted into a plasmid and transfected into separate H uh7.5 human hepatoma cells stably transfected with subgenomic replicons for genotypes 2a, 3a and 4a. All replicons included a firefly luciferase reporter gene. After 48 h of plasmid transfection, the inhibition of HCV replication was determined by HCV RNA isolation and quantification by real‐time polymerase chain reaction and luciferase assays. Results All the secondary structural analogues to genotype 1b X region cross‐inhibited genotype 2a, 3a and 4a replicons. The maximum inhibition by genotype 1b X region structural analogues was obtained against genotype 2a cells in which X 12, X 12a and X 12b inhibited replication by 30%, 63% and 72%, respectively ( P < 0.05 for all), compared to an unrelated hepatitis B viral analogue. Conclusions Despite substantial sequence dissimilarity, HCV RNA genotype 1b X region analogues cross‐inhibited the replication of HCV genotypes 2a, 3a and 4a. Particular conformations and not the sequence of the stem‐loops of the X region are involved in HCV replication.