Prss8^em1Bug

A 2 kb donor or template DNA sequence, containing the desired GC-to-AG mutation to change arginine 44 to glutamine (p.R44Q) in exon 3, was used with ZNF nuclease 5'-TGCCGTCATCCAGCCACGCATCACCGGTGGTGGCAGTG-3' to create this zymogen-locked allele. The point mutation prevents the inactive zymogen pro-form from being activated through proteolitic cleavage into the catalytically active protease form. RT-PCR, Western blots and immunohistochemistry showed that transcription, protein expression and protein location are not affected. (J:231031)