CRISPR/Cas9-mediated genome-edited mice reveal 10 testis-enriched genes are dispensable for male fecundity
CRISPR/Cas9-mediated genome-edited mice reveal 10 testis-enriched genes are dispensable for male fecundity
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DOI：
10.1093/biolre/ioaa084
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发表时间：
2020-08-01
影响因子：
3.6
通讯作者：
Ikawa, Masahito
中科院分区：
文献类型：
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作者：
Park, Soojin;Shimada, Keisuke;Ikawa, Masahito
随着世界人口继续增加到不可持续的水平，控制生育和开发新的避孕药具的重要性正在显现。到目前为止，男性避孕选择一直落后于女性，而且这几种选择并不是每个人都满意的。为了解决这个问题，我们一直在寻找新的候选靶蛋白的非激素避孕药。睾丸特异性蛋白是男性避孕药的有吸引力的靶点，因为它们更可能参与男性生殖，并且预计小分子对它们的靶向作用对其他器官没有靶向有害影响。使用计算机分析，我们确定Erich 2，Glt 6d 1，Prss 58，Slfn 11，Spp 12 c，Stpg 3，Tex 33和Tex 36为小鼠和人类睾丸丰富的基因。基因4930402 F06 Rik和4930568 D16 Rik是睾丸丰富的Glt 6d 1旁系同源基因，我们也在小鼠中发现，但在人类中没有发现，并且也被纳入我们的研究以消除潜在的补偿。我们使用CRISPR/Cas9系统产生了所有列出的基因的敲除（KO）小鼠系。对所有单个KO小鼠系以及Glt 6d 1/4930402 F06 Rik/4930568 D16 Rik TKO小鼠系的分析揭示，它们是雄性可育的，在生殖器官中没有可观察到的缺陷，这表明这10个基因不是雄性可育性所需的，在3个GIt 6D 1旁系同源物的情况下也不起多余的作用。需要进一步的研究来揭示蛋白质的功能，但使用CRISPR/Cas9系统进行体内功能筛选是一种快速准确的方法来寻找男性生育力所必需的基因，这可能适用于其他地方表达的基因的研究。在这项研究中，虽然我们找不到任何潜在的非激素男性避孕药的蛋白质靶点，但我们的发现有助于简化寻找和专注于必需基因的工作。
As the world population continues to increase to unsustainable levels, the importance of birth control and the development of new contraceptives are emerging. To date, male contraceptive options have been lagging behind those available to women, and those few options available are not satisfactory to everyone. To solve this problem, we have been searching for new candidate target proteins for non-hormonal contraceptives. Testis-specific proteins are appealing targets for male contraceptives because they are more likely to be involved in male reproduction and their targeting by small molecules is predicted to have no on-target harmful effects on other organs. Using in silico analysis, we identified Erich2, Glt6d1, Prss58, Slfn11, Spp12c, Stpg3, Tex33, and Tex36 as testis-abundant genes in both mouse and human. The genes, 4930402F06Rik and 4930568D16Rik, are testis-abundant paralogs of Glt6d1 that we also discovered in mice but not in human, and were also included in our studies to eliminate the potential compensation. We generated knockout (KO) mouse lines of all listed genes using the CRISPR/Cas9 system. Analysis of all of the individual KO mouse lines as well as Glt6d1/4930402F06Rik/4930568D16Rik TKO mouse lines revealed that they are male fertile with no observable defects in reproductive organs, suggesting that these 10 genes are not required for male fertility nor play redundant roles in the case of the 3 GIt6D1 paralogs. Further studies are needed to uncover protein function(s), but in vivo functional screening using the CRISPR/Cas9 system is a fast and accurate way to find genes essential for male fertility, which may apply to studies of genes expressed elsewhere. In this study, although we could not find any potential protein targets for non-hormonal male contraceptives, our findings help to streamline efforts to find and focus on only the essential genes.