Genetic Resistance to Avian Leukosis Viruses Induced by CRISPR/Cas9 Editing of Specific Receptor Genes in Chicken Cells

• Published in: Viruses Vol. 10; no. 11; p. 605
• Main Authors: Koslová, Anna, Kučerová, Dana, Reinišová, Markéta, Geryk, Josef, Trefil, Pavel, Hejnar, Jiří
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 02-11-2018; MDPI
• Subjects: Animals; Avian leukosis; Avian Leukosis - genetics; Avian Leukosis - virology; avian leukosis virus; Avian Leukosis Virus - physiology; Base Sequence; Cell Line; Chickens; Codons; CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Disease Resistance - genetics; Gene Editing; Genes, Viral; Genetic Techniques; Genetic Vectors - genetics; Genome editing; Genomes; Inbreeding; Kinases; Leukosis; Mutation; Phenotypes; Poultry; Receptors, Virus - genetics; Receptors, Virus - metabolism; retrovirus receptor; RNA, Guide, CRISPR-Cas Systems; virus-resistance in chicken; Viruses; England; St Louis Missouri; United Kingdom > UK; United States > US; avian leukosis virus; CRISPR/Cas9; virus-resistance in chicken; retrovirus receptor
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v10110605

Avian leukosis viruses (ALVs), which are pathogens of concern in domestic poultry, utilize specific receptor proteins for cell entry that are both necessary and sufficient for host susceptibility to a given ALV subgroup. This unequivocal relationship offers receptors as suitable targets of selection and biotechnological manipulation with the aim of obtaining virus-resistant poultry. This approach is further supported by the existence of natural knock-outs of receptor genes that segregate in inbred lines of chickens. We used CRISPR/Cas9 genome editing tools to introduce frame-shifting indel mutations into
,
, and
loci encoding receptors for the A, C, and J ALV subgroups, respectively. For all three loci, the homozygous frame-shifting indels generating premature stop codons induced phenotypes which were fully resistant to the virus of respective subgroup. In the
locus, we also obtained in-frame deletions corroborating the importance of W38 and the four amino-acids preceding it. We demonstrate that CRISPR/Cas9-mediated knock-out or the fine editing of ALV receptor genes might be the first step in the development of virus-resistant chickens.