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

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 crit...

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Published in:	Proceedings of the National Academy of Sciences - PNAS Vol. 117; no. 4; pp. 2108 - 2112
Main Authors:	Koslová, Anna, Trefil, Pavel, Mucksová, Jitka, Reinišová, Markéta, Plachýa, Jiří, Kalinab, Jiří, Kučerová, Dana, Geryk, Josef, Krchlíková, Veronika, Lejčková, Barbora, Hejnar, Jiří
Format:	Journal Article
Language:	English
Published:	United States National Academy of Sciences 28-01-2020
Subjects:	Amino acids Avian leukosis Biological Sciences Birds Chickens Clonal deletion CRISPR Deletion Food sources Genetic modification Germ cells Heterozygotes Leukosis Na+/H+-exchanging ATPase Nhe1 gene Poultry Subgroups Tryptophan Viruses primordial germ cells CRISPR/Cas9 genome editing in chicken disease resilience in poultry avian leukosis virus subgroup J Na+/H+ exchanger type 1
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Summary:	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 were ALV-J–susceptible. Deletion of W38 did not manifest any visible side effect. Our data clearly demonstrate the antiviral resistance conferred by precise CRISPR/Cas9 gene editing in the chicken. Furthermore, our highly efficient CRISPR/Cas9 gene editing in primordial germ cells represents a substantial addition to genotechnology in the chicken, an important food source and research model.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 1Present address: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, 69120 Heidelberg, Germany. Edited by Stephen H. Hughes, NIH, National Cancer Institute, HIV Drug Resistance Program, Frederick, MD, and accepted by Editorial Board Member Stephen P. Goff December 18, 2019 (received for review August 10, 2019) Author contributions: A.K., P.T., and J.H. designed research; A.K., P.T., J.M., M.R., J.P., J.K., D.K., J.G., V.K., B.L., and J.H. performed research; A.K., P.T., J.M., M.R., J.P., J.K., D.K., J.G., V.K., and J.H. analyzed data; and A.K., P.T., J.M., J.K., and J.H. wrote the paper.
ISSN:	0027-8424 1091-6490
DOI:	10.1073/pnas.1913827117