Precise CRISPR/Cas9 editing of the NHE1 gene renders chickens resistant to the J subgroup of avian leukosis virus

作者：Anna Koslová, P. Trefil, J. Mucksová, M. Reinišová, J. Plachy, J. Kalina, D. Kučerová, J. Geryk, Veronika Krchlíková, Barbora Lejčková, J. Hejnar · 发表于：Proceedings of the National Academy of Sciences of the United States of America · 年份：2020 · DOI：10.1073/pnas.1913827117 · 被引用次数：93 · 研究领域：Biology、Medicine

Significance The current progress of genome editing techniques accelerates the knock-out and knock-in studies in animal models and production of genetic modifications in livestock. Increased resistance to viral pathogens is a particular goal because many monogenic host cell factors are necessary for productive infection and pathogenesis. For example, virus receptors with their specific virus binding sites are direct targets for the CRISPR/Cas9 gene editing. We introduced a single amino acid deletion into the gene encoding the receptor that is required for avian leukosis virus subgroup J to infect chicken cells. Here, we demonstrate that this mutation confers the resistance of chickens to avian leukosis virus subgroup J, an important pathogen in poultry. In addition, we present highly efficient genome-editing technology in chicken. Avian leukosis virus subgroup J (ALV-J) is an important concern for the poultry industry. Replication of ALV-J depends on a functional cellular receptor, the chicken Na+/H+ exchanger type 1 (chNHE1). Tryptophan residue number 38 of chNHE1 (W38) in the extracellular portion of this molecule is a critical amino acid for virus entry. We describe a CRISPR/Cas9-mediated deletion of W38 in chicken primordial germ cells and the successful production of the gene-edited birds. The resistance to ALV-J was examined both in vitro and in vivo, and the ΔW38 homozygous chickens tested ALV-J–resistant, in contrast to ΔW38 heterozygotes and wild-type birds, which we...