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. 2023 Feb 14;120(8):e2207263120. doi: 10.1073/pnas.2207263120

Fig. 2.

Fig. 2.

1700029I15Rik Knockout Males Are Infertile despite Normal Spermatogenesis and Sperm Morphology. (A) CRISPR/Cas9-mediated knockout of 1700029I15Rik in the B6D2F1 hybrid mouse strain. Single guide RNAs (sgRNAs) #1 and #2 were designed to target the first coding exon (Exon 1) and the 3′ untranslated region (UTR), respectively. Forward (Fw) and reverse (Rv) primers flanking the deletion region were employed for genotyping. (B) Genomic PCR for detecting wild-type and 1700029I15Rik knockout alleles. (C) Western blot detection of 1700029I15Rik in wild-type and knockout testes and sperm. BASIGIN was analyzed in parallel as a loading control. (D) Fertility tests of wild-type, 1700029I15Rik–/–, and 1700029I15Rik–/–; Tg male mice. ns, not significant. (E) Fertility tests of wild-type, 1700029I15Rik+/Δ, and 1700029I15RikΔ/Δ mice. The mutant mouse line was produced on an inbred genetic background of C57BL/6J. (F) Analysis of testis weight in wild-type and 1700029I15Rik–/– males. (G) Analyses of testis histology and sperm morphology in 1700029I15Rik+/– and 1700029I15Rik–/– males.