H1N1 SIV replication is increased in Plscr1 knockout
mice
In order to investigate the functions of PLSCR1 during Influenza A virus
infection in vivo , we developed Plscr1 knockout mice using
CRISPR/Cas9 genome editing technique. The mouse Plscr1 gene
contains eight exons encoding 378 amino acids, and two sgRNAs were
designed to target exon 4 (Figure 2A). DNA sequencing revealed that a
deletion of 122 bp was introduced into exon 4 in Plscr1 knockout
mice, which causes a premature translational stop and gives rise to a
potentially truncated protein of 43 amino acids (Figure 2B). RT-PCR,
western blot, and immunohistochemistry results confirm that PLSCR1
expression is indeed disrupted in the homogenous knockout mice (Figures
2C-2E). Interbreeding of Plscr1+/− mice gave
rise to offspring in the expected Mendelian ratio (23.8% wild-type,
47.6% Plscr1+/− , and 28.6%Plscr1-/- ; n=42) with normal viability,
suggesting that PLSCR1 is dispensable for general development in mice.