Transgenic mice expressing a soluble form of porcine nectin-1/herpesvirus entry mediator C as a model for pseudorabies-resistant livestock

Abstract

An approach to genetically engineered resistance to pseudorabies virus (PRV) infection was examined by using a transgene encoding a soluble form of nectin-1, also known as herpesvirus entry mediator C. Nectin-1 is an ␣-herpesvirus receptor that binds to virion glycoprotein D. Nectin-1 mediates entry of PRV, herpes simplex virus types 1 and 2, and bovine herpesvirus type 1. To assess the antiviral potential of an ectopic expression of the nectin-1 ectodomain in vivo, six transgenic mouse lines expressing a soluble form of nectin-1, consisting of an extracellular domain of porcine nectin-1 and the Fc portion of human IgG1, were generated. All of the transgenic mouse lines showed nearly complete resistance to PRV infection by means of both i.p. and intranasal routes. These results suggest that the introduction into farm animals of a transgene encoding a soluble form of nectin-1 would offer a potent biological approach to generating ␣-herpesvirusresistant livestock.

Microphthalmia and lack of vitreous body in transgenic mice expressing the first immunoglobulin-like domain of nectin-1

“…Transgenic founders were identified by PCR using genomic DNA isolated from their tails as described previously [9]. Presence of PHveC-VhIg in sera of transgenic mice was confirmed by Western blot analysis as described previously [9]. The transgenic founders were crossed with C57BL/6 mice and their offspring were used for the experiments.…”

“…The transgenic founders were crossed with C57BL/6 mice and their offspring were used for the experiments. The transgenic mouse lines PHveCIg6, 22, 32, 33 and 37 expressing PHveCIg made of the entire ectodomain of nectin-1 fused to human Fc portion [9], PHveCpIg150, 151 and 185 expressing PHveCpIg made of the entire ectodomain of nectin-1 fused to porcine Fc portion [11], and PHveC-VpIg21, 30 and 72 expressing PHveC-VpIg made of the first Ig-like domain of nectin-1 fused to porcine Fc portion [12] were used for experiments. Schematic representation of each fusion protein is shown in Fig.…”

“…1a) containing the CAG promoter [7] (cytomegalovirus IE enhancer and chicken β-actin promoter), the PHveC-VhIg gene and the rabbit β-globin polyA signal was microinjected into C57BL/6 mouse eggs to generate transgenic mice. Transgenic founders were identified by PCR using genomic DNA isolated from their tails as described previously [9]. Presence of PHveC-VhIg in sera of transgenic mice was confirmed by Western blot analysis as described previously [9].…”

Transgenic Modification of Production Traits in Farm Animals

“…Transgenic mice expressing a soluble form of a protein known as herpesvirus entry mediator were generated. The transgenic mice had nearly complete resistance to challenge with the alpha-herpesvirus, suggesting the potential of the approach for generating pseudorabies-resistant livestock [149].…”

Relations between animal transgenesis and reproduction

“…The virus is thus trapped by the overexpressed soluble receptor and becomes unable to infect cells and animals. Generation of transgenic pigs resistant to Aujeszky disease is now conceivable [32].…”