Creation of a Recombinant Adenovirus to Observe Infection in Real Time
Additional Funding Sources
The project described was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant No. P20GM103408.
Presentation Date
7-2020
Abstract
Adenovirus 14p1 (Ad14p1) is an emerging strain of adenovirus 14 (Ad14) that induces a more severe response in otherwise healthy patients. Infection of Syrian hamsters with Ad14p1 results in bi-lateral patchy bronchopneumonia that is not seen with Ad14 infection. The reason for this increased pathogenicity between Ad14 and Ad14p1 is unknown. The E3 region of the Ad genome is not required for viral replication and is often deleted to allow for expression of various proteins. The objective of my research project was to construct an E3 deleted Ad14 and Ad14p1 and insert a Red Fluorescent Protein-luciferase (RFP-luc) cassette to allow for real time observation of viral infection in hamsters. The E3 deletion (ΔE3) was introduced into the Ad14 and Ad14p1 clones by homologous recombination and Ad14ΔE3 and Ad14p1ΔE3 viruses were recovered in A549 cells. Viral fitness of the ΔE3 viruses was assessed by plaque assay and one-step growth curves. The RFP-luciferase construct was introduced in Ad14 and Ad14p1 by homologous recombination and viruses are currently being recovered. Our results suggest that the ΔE3 viruses can be used as a genetic tool to look at both viral and host factors that determine Ad14p1 pathogenesis.
Creation of a Recombinant Adenovirus to Observe Infection in Real Time
Adenovirus 14p1 (Ad14p1) is an emerging strain of adenovirus 14 (Ad14) that induces a more severe response in otherwise healthy patients. Infection of Syrian hamsters with Ad14p1 results in bi-lateral patchy bronchopneumonia that is not seen with Ad14 infection. The reason for this increased pathogenicity between Ad14 and Ad14p1 is unknown. The E3 region of the Ad genome is not required for viral replication and is often deleted to allow for expression of various proteins. The objective of my research project was to construct an E3 deleted Ad14 and Ad14p1 and insert a Red Fluorescent Protein-luciferase (RFP-luc) cassette to allow for real time observation of viral infection in hamsters. The E3 deletion (ΔE3) was introduced into the Ad14 and Ad14p1 clones by homologous recombination and Ad14ΔE3 and Ad14p1ΔE3 viruses were recovered in A549 cells. Viral fitness of the ΔE3 viruses was assessed by plaque assay and one-step growth curves. The RFP-luciferase construct was introduced in Ad14 and Ad14p1 by homologous recombination and viruses are currently being recovered. Our results suggest that the ΔE3 viruses can be used as a genetic tool to look at both viral and host factors that determine Ad14p1 pathogenesis.