Identification of TTC29 protein partners in mouse testis

We previously identified gene variants in TTC29 that induce human male infertility due to asthenozoospermia and MMAF phenotype (Lores et al. Am J hum genet 2019). By means of CRISPR/Cas9 mutagenesis we demonstrated that the orthologous TTC29 protein in mouse also constitute a flagellar protein that is required for proper beating of the flagella, indicating an evolutionary-conserved function of TTC29. In here, we took advantage of the mutant mouse model to further decipher the protein complexes associated with TTC29 function within the flagella. We performed co-immunoprecipitation experiments on testis protein extracts from wild-type mice using TTC29 specific antibodies and performed comparative mass spectrometry analyses with two TTC29 KO mouse lines we previously generated and shown to lack TTC29 protein (Ttc29-/- L5 and L7) (Lores et al. Am J hum genet 2019. Mass spectrometry analyses were performed on four independent experimental replicates for each mouse line.