| Introduction: Hepatitis C virus (HCV) is a blood-borne pathogen, resulting in liver cirrhosis and liver cancer.
Despite of many efforts in development of treatments for HCV, no vaccine has been licensed yet. The purpose of
this study was to design and prepare a specific mRNA, without 5' cap and poly (A) tail transcribed in vitro capable
of coding core protein and also to determine its functionality. Methods: Candidate mRNA was prepared by in vitro
transcription of the designed construct consisting of 5ʹ and 3ʹ untranslated regions of heat shock proteins 70
(hsp70) mRNA, T7 promoter, internal ribosome entry site (IRES) sequences of eIF4G related to human dendritic
cells (DCs), and the Core gene of HCV. To design the modified mRNA, the 5' cap and poly (A) tail structure was not
considered. DCs were transfected by in vitro-transcribed messenger RNA (IVT-mRNA) and the expressions of
green fluorescent protein (GFP), and Core genes were determined by microscopic examination and Western
blotting assay. Results: Cell transfection results showed that despite the absence of 5' cap and poly (A) tail, the
structure of the mRNA was stable. Moreover, the successful expressions of GFP and Core genes were achieved.
Conclusion: Our findings indicated the effectiveness of a designed IVT-mRNA harboring the Core gene of HCV in
transfecting and expressing the antigens in DCs. Considering the simple and efficient protocol for the preparation
of this IVT-mRNA and its effectiveness in expressing the gene that it carries, this IVT-mRNA could be suitable for
development of an RNA vaccine against HCV. |