Skip to main content
. 2020 Apr 17;103(2):244–253. doi: 10.1093/biolre/ioaa040

Figure 2.

Figure 2

Generating Ssmem1 KO mice. (A) Genomic structure of mouse Ssmem1 and scheme to generate the gene KO mice using the CRISPR/Cas9 system. Three splice variants of Ssmem1 have been reported. White and black boxes indicate untranslated and coding regions, respectively. Red arrow and black under bar indicate gRNA that targets the region. The delivery of CRISPR/Cas9 system into zygotes for mutagenesis via microinjection resulted in a 5 bp (6 bp deletion and 1 bp insertion) frameshift deletion (shown in red). (B) Genotyping of Ssmem1 alleles. Primers indicated in green color in (A) amplify specific amplicon for the WT or KO allele. (C) Mutation in amino acid sequence in KO mouse induced by—five frameshift mutation is shown in red color. Asterisk indicates a premature stop codon. The numbers above sequences are amino acid number for each. (D) Western blot analysis using testis and sperm. SSMEM1 protein is detected only in WT testis. Equal loading of total protein was confirmed by Coomassie Brilliant Blue staining.