Knock-Out of Retrovirus Receptor Gene <i>Tva</i> in the Chicken Confers Resistance to Avian Leukosis Virus Subgroups A and K and Affects Cobalamin (Vitamin B<sub>12</sub>)-Dependent Level of Methylmalonic Acid
2021
Anna Koslová | Pavel Trefil | Jitka Mucksová | Veronika Krchlíková | Jiří Plachý | Jakub Krijt | Markéta Reinišová | Dana Kučerová | Josef Geryk | Jiří Kalina | Filip Šenigl | Daniel Elleder | Viktor Kožich | Jiří Hejnar
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 <i>tva</i> locus with the aim of knocking-out Tva expression and creating a virus-resistant chicken line. The <i>tva</i> 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 <i>tva</i> −/− 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 <i>tva</i> +/+ and <i>tva</i> +/− siblings. We also found a specific disorder of the cobalamin/vitamin B<sub>12</sub> 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.
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