Generation and functional characterization of a conditional Pumilio2 null allele

摘要

The highly conserved RNA binding protein PUF (Pumilio/FBF) family is present throughout eukaryotes from yeast to mammals,with critical roles in development,fertility and the nervous system.However,the function of the mammalian PUF family members remains underexplored.Our previous study reported that a gene-trap mutation of Pum2 results in a smaller testis but does not impact fertility and viability.Although the gene-trap mutation disrupted the key functional domain of PUM protein-PUM-HD (Pumilio homology domain),but still produced a chimeric Pum2-β-geo protein containing part of PUM2,raising a question if such a chimeric protein may provide any residual function or contribute to the reproductive phenotype.Here,we report the generation of a conditional PUM2 allele,when knocked out,producing no residual PUM2 and hence a complete loss-of-function allele.We also uncovered small but significant reduction of male fertility and viability in the mutants,suggesting requirement of PUM2 for male fertility and viability.