Development and characterization of a synthetic infectious cDNA clone of the virulent Bucyrus strain of equine arteritis virus expressing mCherry (red fluorescent protein).

This research discusses the creation and examination of a synthetic infectious cDNA clone of the virulent Bucyrus strain of equine arteritis virus (EAV), coded with mCherry, a red fluorescent protein. This study highlights the value of using advances in nucleic acid synthesis technology as a research tool for identifying how a virus infects different cell types and how these viral proteins attach and enter various cell subpopulations.

Creation and Examination of a Synthetic Infectious cDNA Clone of EAV

• The researchers designed and synthesised an infectious cDNA clone of the horse-adapted virulent Bucyrus strain (VBS) of EAV, which was also encoded with mCherry, a red fluorescent protein. This was done using advances in in silico design and de novo nucleic acid synthesis technology.
• The resulting progeny virions (EAV sVBSmCherry) were then characterized in vitro based on their ability to infect cells, their replicative capacity, and the stability of the mCherry coding sequences after repeated passage in cell culture.

Use of mCherry Sequence as a Research Tool

• The study found that the mCherry sequence remained relatively stable during sequential cell culture passage, suggesting that the EAV-sVBSmCherry-derived virus could serve as a valuable research tool.
• This tool could be used to identify which cell types are prone to infection by the virus (host-cell tropism) and to characterize the proteins in the virus that facilitate its attachment and entry into various subpopulations of peripheral blood mononuclear cells.

Advantages of Nucleic Acid Synthesis Technology

• The study also shows that using the latest methods in nucleic acid synthesis allows for the creation of complex viral genomes, such as those found in arteriviruses, without resorting to the complex techniques traditionally used in molecular cloning.
• This technology makes the process of designing viral vectors more efficient and less risk-ridden, bypassing the difficulties and risks associated with conventional lab techniques.