Influenza A virus genome segment 7 encodes protein M1, which is the matrix protein playing crucial role in the virus life cycle. Any antiviral strategy that aims at reducing, in particular, the expression of this genome segment should, in principle, reduce the infectivity of the virus. We developed a specific antiviral approach at the molecular level and designed several novel 10-23 DNAzymes (Dz) and hammerhead ribozymes (Rz), specifically targeted to cleave at the conserved domains of the influenza virus M1 RNA. We sought to use antisense molecules with the hope that it will facilitate the ribozyme-mediated cleavage. We observed that the Mg 2+-dependent sequence-specific cleavage of M1 RNA was achieved by both the Dz and Rz in a dose-dependent manner. This combination of catalytic Dz and Rz with antisense molecules, in principle, resulted in more effective gene suppression, inhibited the whole virus replication in host cell, and thus could be exploited for therapeutic purposes.
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Fig. 9 The MDCK cells were transfected with 3 µg each of ribozyme and DNAzyme individually as well as synergistically and subjected to virus challenge (MOI of 0.1) at 24 h post-transfection. Total RNA was isolated and subjected to real-time PCR analysis to detect the fold change in expression of M1 gene of influenza A virus Acknowledgments The authors thankfully acknowledges the financial support provided by Department of Biotechnology, Government of India.
- Gene silencing
- Influenza virus