Location: Foreign Animal Disease Research
Project Number: 8064-32000-060-16-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Apr 1, 2018
End Date: Nov 30, 2020
Objective:
This collaborative research project focuses in the use of Porcine Cytomegalovirus (pCMV) as an immunogenic vector to deliver recombinant antigens against Classical Swine Fever Virus (CSFV) strain E2 and Porcine Reproduction and Respiratory Syndrome (PRRS) virus antigens.
The objectives of this project are:
1. Evaluate and select one or more avirulent Korean pCMV isolates based on swine safety and immune evasion characterization studies.
2. Establish pCMV gene editing and recombinant pCMV construction techniques.
3. Construct and characterize two recombinant pCMVs; a) PRRS Virus protective T cell antigen and b) Expressing CSFV protective B cell antigen.
Approach:
Naturally attenuated field isolates of pCMV will be characterized to determine their level of virulence in pathogenesis studies. Those isolates proven to be attenuated, although still able to mount a specific response, will be genetically characterized by full-length sequence using next generation sequencing (NGS). The information obtained will be used to develop a methodology, based in CRISP/Cas9 approach, to produce recombinant pCMV. Two different recombinant pCMV will be created harboring recombinant genes; the CSFV major structural glycoprotein E2 and the major T cell epitope from PRRS virus. Recombinant viruses will be evaluated in terms of recombinant protein expression and genetic stability.
