Location: Foreign Animal Disease Research
Project Number: 3022-32000-063-011-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2019
End Date: May 15, 2024
Objective:
This research project will aid in the development of control strategies against African Swine Fever Virus (ASFV), specifically;
(i) determining what protein epitopes in ASFV are serologically recognized by pigs and developing antibodies and ELISAs to these protein epitopes, and
(ii) development of stable cells from domestic, or wild pigs and or ticks, supporting the replication of those vaccine candidates
Approach:
The two objectives of this project will be pursued separately in methodological
terms.
(i) ELISAs will be developed to specifically detect the presence of antibody responses in pigs to the ASFV proteins that will then be expressed and further purified as recombinant proteins using the baculovirus as expression system. Those proteins will be then used to develop and optimize direct ELISAs which will be used to detect the presence or absence of specific antibodies in vaccinated and ASFV infected pigs.
(ii) Host cells will be derived from ASFV natural hosts, ie. domestic and wild swine, as well as ticks, as potential sources to establish a cell line supporting virus replication. Cells will be immortalized by different methodologies, and the surviving cell lines cloned to establish stable cell clones. Each of those established cell lines will be tested in their ability to support virus replication. Those selected as the better substrate for virus production will be evaluated to ensure they are free of adventitious contaminants.
iii) CRISPR sgRNA Library screening with a nuclease deficient Cas9 for gene silencing in cell lines permissive to ASFV towards the identification of genes cells which support ASFV replication. This will determine potential genetic targets that could be used to make geneticially modified cells resistant or permissive to infection.
