Optimization and Stability studies of siRNA for Significant Inhibition of Dengue (DENV-2) Viral Replication in Vero cells

Order of Publishing in Issue: 
1
Volume :10
Issue :1
January, 2016
Page No: 
13-19
Authors: 
Anil Babu Korrapati[1,2]* and Muvva Vijayalakshmi[2]
Address: 
[1] Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India
Address: 
[2] Department of Botany and Microbiology, Acharya Nagarjuna University, Nagarjuna Nagar-522510, Guntur, Andhra Pradesh, India.

RNA interference (RNAi) is a highly conserved, specific and selective biological response of gene regulation mechanism and has been widely exploited for designing novel drug targets to treat infections caused by pathogens including viruses. In the previous study, one of the promising sh-5b RNA (short hairpin RNA, sh- 5b) construct was targeted to the conserved sequence within the 5¹-NTRs (non-translated regions) of all Dengue serotypes. The current study report the stability (up to 24 months) of recombinant adenovirus (rAd) engineered to express sh-5b RNA for DENV-2 replication inhibition. Stability study was done by gene specific PCR, where as the effect of sh-5b RNA in attenuating DENV-2 replication was further validated by One step RT-PCR by TITAN. The rAds harboring sh-5b RNA to target dengue serotypes might serve as an efficient alternative to conventional drugs for therapeutic intervention to treat DENV infections in humans.

Keywords: 
RNA interference, Dengue, short hairpin RNA, non-translated region, One step RTPCR, gene specific PCR
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