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Tanapoxvirus (TPV), a poxvirus originally isolated from the Tana River Valley in equatorial Africa (Downie & España, 1972), appears to be an ideal oncolytic virus (OV). Some characteristics that make it an appealing OV include a large viral genome, thermostability, and its inability to spread from human to human. TPV causes a mild self-limiting infection in humans and monkeys and is epidemiologically restricted in equatorial Africa. The rest of the global population is immunologically naïve to TPV (Zhang et al., 2018). The role of IL-2 in T-cell and other immune cell proliferation has previously been described (Gaffen & Liu, 2004), and IL-2 gene insertion into an OV vector has demonstrated promising results in regressing human tumors (Suryawanshi et al., 2017; Zhang et al., 2018). Thymidine kinase (TK) is an enzyme necessary for viral DNA replication and is over-expressed in tumor cells (Hwang et al., 2011). Therefore, removal of viral TK gene from the viral genome makes cytoplasmic DNA viruses preferentially replicate in tumor cells.
We have employed homologous recombination to insert human IL-2 (hIL-2) gene into a p2KO plasmid vector (Conrad et al., 2015), and then into the TPV viral genome by transfecting it into TPV-infected cells. The hIL-2 insertion ablated TPV-66R, which encodes viral TK. PCR was used to confirm the presence of IL-2 and deletion of TK in the viral genome.
Woyczesczyk, Helen, "Insertion of IL-2 Gene into Tanapoxvirus Genome" (2018). Honors Theses. 3060.
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