Life Sciences & Biotechnology
Title : | Application of biofunctionalized nanoparticles to delivery targeted siRNA in shrimp model to control White Spot Syndrome Virus |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Anusha J.R., Centre For Marine Science And Technology (CMST), Uttar Pradesh |
Timeline Start Year : | 2023 |
Timeline End Year : | 2025 |
Contact info : | anushajr@gmail.com |
Details
Executive Summary : | White Spot Syndrome Virus (WSSV) is a problematic pathogen, which causes White Spot Disease (WSD) in Crustaceans especially in shrimps. Various preventive measures such as disinfection, use of WSSV resistant strains and other bio-secure techniques were practiced. However, these preventive strategies cannot be effective once the shrimp farm is infected. There is no effective commercial product to protect against the disease in shrimp. The preventive and treatment strategy for viral infections are very challenging, because it uses the host cell to replicate. Hence, there is an immediate need to develop a novel therapeutic vaccine that can ensure effective protection from WSSV in shrimp. Objective: The major objective is to develop a dual-purpose, targeted viral protein biofunctionalized with therapeutic metal nanoparticle vaccine for WSD in shrimp, Litopenaeus vannamei. Major experiments: The envelope protein, VP19 which is a DNA binding protein in WSSV will be selected for developing vaccine. The VP19-siRNA with restriction enzymes were annealed and cloned into the Baculovirus vector. The existence of recombinant plasmid will be confirmed by DNA sequencing and transferred to E. coli. In addition, the expression of VP19-siRNA in E. coli will be confirmed using Northern blotting. To overcome the barriers associated with siRNA trafficking mechanism, metal-chitosan nanoparticles will be incorporated. For that, the silver-chitosan nanoparticles (Ag/CSNPs) will be synthesized, characterized and the antiviral efficiency will be examined in WSSV infected shrimp. The Ag/CSNPs biofunctionalized with siRNA-VP19 will be injected in shrimp and infected with WSSV to analyze vaccine potential. The gene expression will be studied using Western blotting and qRT-PCR. The immunological response, immunohistochemical parameters and histopathological studies will be performed and statistically analyzed using the SPSS software. This new approach, siRNA-VP19 biofunctionalized Ag/CSNPs drug/vaccine will surely enhance and improve the targeted immune response. The envelope protein, VP19 will enhance the survivability of shrimp after WSSV infection. The siRNA incorporation will demonstrate a significant antiviral performance, and suggest an efficient approach. The Ag/CSNPs will acts as anti-viral drug; enhance the stability of the vaccine; easily pass through transmembrane without much transfection agents; protect the therapeutic drugs from degradation; easy drug intake and enhance the therapeutic potential to cure the disease. Success in this project would provide dual purpose drug/vaccine in one shot through a biocompatible source with cost effective and less time-consuming production. The expected outcome of the project would be production of siRNA against WSSV of shrimp and delivery systems using biofunctionalized nanoparticles. These products would be used to develop resistance against viral pathogen in shrimp. |
Organizations involved