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

Proc Natl Acad Sci U S A. 2020 Jan 28;117(4):2108-2112. doi: 10.1073/pnas.1913827117. Epub 2020 Jan 21.

Abstract

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.

Keywords: CRISPR/Cas9 genome editing in chicken; Na+/H+ exchanger type 1; avian leukosis virus subgroup J; disease resilience in poultry; primordial germ cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Animals, Genetically Modified / immunology
  • Animals, Genetically Modified / virology
  • Avian Leukosis / genetics
  • Avian Leukosis / immunology*
  • Avian Leukosis / virology
  • Avian Leukosis Virus / classification
  • Avian Leukosis Virus / genetics*
  • Avian Leukosis Virus / physiology
  • Avian Proteins / genetics*
  • Avian Proteins / immunology
  • CRISPR-Cas Systems
  • Chickens
  • Disease Resistance
  • Female
  • Gene Editing
  • Male
  • Poultry Diseases / genetics
  • Poultry Diseases / immunology*
  • Poultry Diseases / virology
  • Sodium-Hydrogen Exchanger 1 / genetics*
  • Sodium-Hydrogen Exchanger 1 / immunology

Substances

  • Avian Proteins
  • Sodium-Hydrogen Exchanger 1