en بيولوژي Biology پژوهشي Research <div style="text-align: justify;"><strong><span dir="LTR">Background: </span></strong><span dir="LTR">Spermatogenesis associated 19 (<em>Spata</em>19) was introduced as a testis-specific gene</span><span dir="LTR"> that was probably involved in spermatogenesis cell apoptosis. Therefore, this study aimed to investigate the effect of <em>Spata</em>19 inactivation on sperm count.</span><br> <strong><span dir="LTR">Materials and Methods: </span></strong><span dir="LTR">We generated global <em>Spata</em>19 knockout mice by CRISPR/Cas9</span><span dir="LTR"> nickase technology. Disability was validated in three levels of DNA, RNA, and protein using</span><span dir="LTR"> PCR, RT-PCR, and immunohistochemistry. Histological studies were performed for testis. Sperm characteristics were also assessed with CASA software.</span><br> <strong><span dir="LTR">Results: </span></strong><em><span dir="LTR">Spata</span></em><span dir="LTR">19 knockout mice had a 43 nucleotides deletion in exon 4 of this gene. The presence</span><span dir="LTR"> and absence of <em>Spata</em>19 were confirmed in normal and knockout mice, respectively. The presence</span><span dir="LTR"> of <em>Spata</em>19 in normal NMRI mice was detected in the brain, heart, and thymus by semi-nested</span><span dir="LTR"> RT-PCR and in Leydig cells by immunohistochemistry. Histological studies revealed a decrease in sperm count in knockout mice. Also, CASA parameters were significantly reduced (P<0.05).</span><br> <strong><span dir="LTR">Conclusion: </span></strong><span dir="LTR">These data indicate that <em>Spata</em>19 inactivation is a cause of oligospermia, and its role</span><span dir="LTR"> could be beyond an adhesive molecule.</span></div> spermatogenesis associated 19 (Spata19), CRISPR/Cas9 nickase, global knockout mice, oligospermia, infertility, spermatogenesis, outbred mice, 29 38 http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1115-2&slc_lang=en&sid=1 Mahsa Zargar mzargar@alumnus.tums.ac.ir 1 10031947532846009448 No Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Abbas Jamshidizad jamshidi_1@yahoo.com 2 10031947532846009449 No Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Aidin Rahim-Tayefeh aidin.tayefe@gmail.com 3 10031947532846009450 No Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Ehsan Hashemi e.hashemy@gmail.com 4 10031947532846009451 No Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Sasan Shabani sshabani@razi.tums.ac.ir 5 10031947532846009452 No Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Mehdi Shamsara Shamsam2000@gmail.com 6 10031947532846009453 No Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Mohammad Hossein Modarressi Modaresi@tums.ac.ir 7 10031947532846009454 Yes Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.