Gene editing of pigs to control influenza A virus infections

• Published in: Emerging microbes & infections 2024-12, Vol.13 (1), p.2387449
• Main Authors: Kwon, Taeyong, Artiaga, Bianca L, McDowell, Chester D, Whitworth, Kristin M, Wells, Kevin D, Prather, Randall S, Delhon, Gustavo, Cigan, Mark, White, Stephen N, Retallick, Jamie, Gaudreault, Natasha N, Morozov, Igor, Richt, Juergen A
• Format: Article
• Language: English
• Subjects: Gene editing; Hogs; Influenza; influenza A virus; knockout pigs; TMPRRS2; transmembrane serine protease 2; Viruses
• ISSN: 2222-1751; 2222-1751
• DOI: 10.1080/22221751.2024.2387449

Proteolytic activation of the hemagglutinin (HA) glycoprotein by host cellular proteases is pivotal for influenza A virus (IAV) infectivity. Highly pathogenic avian influenza viruses possess the multibasic cleavage site of the HA which is cleaved by ubiquitous proteases, such as furin; in contrast, the monobasic HA motif is recognized and activated by trypsin-like proteases, such as the transmembrane serine protease 2 (TMPRSS2). Here, we aimed to determine the effects of TMPRSS2 on the replication of pandemic H1N1 and H3N2 subtype IAVs in the natural host, the pig. The use of the CRISPR/Cas 9 system led to the establishment of homozygous gene edited (GE) knockout (KO) pigs. Delayed IAV replication was demonstrated in primary respiratory cells of KO pigs . IAV infection resulted in significant reduction of virus shedding in the upper respiratory tract, and lower virus titers and pathological lesions in the lower respiratory tract of KO pigs as compared to wild-type pigs. Our findings support the commercial use of GE pigs to mitigate influenza A virus infection in pigs, as an alternative approach to prevent zoonotic influenza A transmissions from pigs to human.