Adenoviral Vector-mediated Editing of chNHE1 Gene for Avian Leukosis Virus Subgroup J Resistance Study
Chicken Na+/H+exchanger type 1(chNHE1)is a cellular receptor for avian leukosis virus subgroup J(ALV-J)and a cellular target of ALV-J infection.To explore the breeding goal of chicken resistance to avian leukaemia,experiment,a targeted CRISPR/Cas9 adenoviral editing system for the chNHE1 gene was constructed and the changes in cellular resistance to ALV-J after editing were evaluated.The results showed that the mutations in chNHE1 were introduced effectively by CRISPR/Cas9 adenoviral vector-mediated homologous recombination protocol of Donor,and there was 36%of monoclonal cell lines with W38 deletion.The cells with T37 deletion possessed highly significant resistance to ALV-J infection.The cells with W38 deletion and the E35A cells with modified P36 and T37 deletions were found to be completely resistant to ALV-J infection.The results of this experiment indicated that chicken DF-1 cells could be edited effectively by CRISPR/Cas9 in adenoviral vector,and either W38 deletion or E35A replacement of P36,T37 deletion could make DF-1 resistant to ALV-J infection.This protocol could be used to a technological stock for breeding chicken lines with resistance to ALV-J infection.
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