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. 2022 Jan 12;9:810118. doi: 10.3389/fcell.2021.810118

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Copyright © 2022 Matsumura, Noda, Satouh, Morohoshi, Yuri, Ogawa, Lu, Isotani and Ikawa.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation of Izumo1-deficient rats using CRISPR/Cas9. (A) Design of sgRNA for disrupting Izumo1 in rats. White and gray boxes represent the untranslated and coding regions, respectively. (B) Genomic Izumo1 sequences of WT and mutant (Izumo1 ^−7/−7) alleles. (C) Detection of rat Izumo1 mRNA in testis by RT-PCR. (D) Immunostaining of IZUMO1 in the testicular sperm from WT or mutant rats. The acrosomal caps and the nuclei were stained by the fluorophore-conjugated Lectin PNA and Hoechst 33342, respectively. Scale bar = 10 µm. (E) The average number of pups per copulatory plug. Three males each were tested for the Izumo1 Het and Izumo1 KO groups. ***p < 0.001.