Journal Article

Knock-Out of Retrovirus Receptor Gene Tva in the Chicken Confers Resistance to Avian Leukosis Virus Subgroups A and K and Affects Cobalamin (Vitamin B12)-Dependent Level of Methylmalonic Acid

Mucksová, Jitka, Krchlíková, Veronika, Geryk, Josef, Hejnar, Jiří, Elleder, Daniel, Reinišová, Markéta, Krijt, Jakub, Kučerová, Dana, Plachý, Jiří, Kožich, Viktor, Šenigl, Filip, Kalina, Jiří, Koslová, Anna, Trefil, Pavel

2021

Overview

The chicken Tva cell surface protein, a member of the low-density lipoprotein receptor family, has been identified as an entry receptor for avian leukosis virus of classic subgroup A and newly emerging subgroup K. Because both viruses represent an important concern for the poultry industry, we introduced a frame-shifting deletion into the chicken tva locus with the aim of knocking-out Tva expression and creating a virus-resistant chicken line. The tva knock-out was prepared by CRISPR/Cas9 gene editing in chicken primordial germ cells and orthotopic transplantation of edited cells into the testes of sterilized recipient roosters. The resulting tva −/− chickens tested fully resistant to avian leukosis virus subgroups A and K, both in in vitro and in vivo assays, in contrast to their susceptible tva +/+ and tva +/− siblings. We also found a specific disorder of the cobalamin/vitamin B12 metabolism in the tva knock-out chickens, which is in accordance with the recently recognized physiological function of Tva as a receptor for cobalamin in complex with transcobalamin transporter. Last but not least, we bring a new example of the de novo resistance created by CRISPR/Cas9 editing of pathogen dependence genes in farm animals and, furthermore, a new example of gene editing in chicken.

• Publisher: MDPI AG,MDPI
• Subject: Animals / Avian leukosis / Avian leukosis virus / Avian Leukosis Virus - classification / Avian Leukosis Virus - physiology / avian leukosis virus subgroups A/K / Avian Proteins - genetics / Avian Proteins - physiology / Birds / Cell surface / Chick Embryo / Chickens - virology / Cloning / CRISPR / CRISPR-Cas systems / Disease resistance / Embryos / farms / Female / Frameshift Mutation / Gamma rays / Gene deletion / Gene Editing / gene editing in chicken / Gene Knockout Techniques / genes / Genome editing / Germ cells / Glycoproteins / Infections / Leukosis / loci / low density lipoprotein / Male / Metabolism / methylmalonic acid / Methylmalonic Acid - blood / Mutation / pathogens / Poultry / poultry industry / Proteins / Receptor density / Receptors, Virus - genetics / Receptors, Virus - physiology / Spermatogenesis / surface proteins / Transplantation / tva / Viruses / Vitamin B 12 - metabolism / Vitamin B12 / vitamin B12/cobalamin