Characterization of non Structural 5A Gene of Hepatitis C Virus (3A Genotype)

Hepatitis C is the commonest among all viral diseases. It is caused by the Hepatitis C Virus. It is a slowly developing blood born disease which severely affects the liver cells and create a lot of medical complications that ultimately leads to hepatocellular carcinoma.3a is the most common genotype in Pakistan. Screening, diagnosing and adequate therapy is a key factor to treat this infection before it gets lethal. NS5A is a viral regulatory protein that modifies viral RNA replication and host processes by cooperating directly and indirectly with a range of host regulatory factors, thus plays a vital role in cellular and viral processes essential for viral life cycle. It can alter cellular response to interferon and association with regulation of interleukin-8 hence, playing an important role in resistance to interferon. Adequate NS5A inhibitors can be a better approach as an antiviral agent by suppression of Hepatitis C virus replication. The present study is an initial step in this regard and will be very helpful in generation of antiviral agents against local HCV type. For this purpose, NS5A gene from 3a genotype was amplified, purified and cloned in E. coli by using TA vector followed by blue/white screening for transformants, colony PCR and restriction digestion. This study can lead to the development of more sensitive, specific and reproducible as compared to commercially available antiviral agents in Pakistan. The in-vitro activity of antiviral agents against purified recombinant protein has laid the foundation to develop an in-house anti-HCV antiviral agent for the most prevalent HCV genotype (3a) in Pakistan. However, development toward this goal will require not only more detailed molecular information but also a vigorous, high-through put assay for selection of compounds able to block NS5A RNA binding.