Generation of a new Cre-deleter mouse line to study spermiogenesis

项目摘要 本提案的目标是生成和表征圆形精子细胞特异性Cre删除小鼠系。 目前，还没有Cre-删除小鼠品系可用于靶向特异性地在哺乳动物中缺失floxed基因。 圆形精子细胞。可用的睾丸生殖细胞Cre-lines删除精原细胞中的floxed基因（Stra 8- iCre）或精母细胞（Spo 11-Cre，Hsp 2a-Cre）中。这些线不适合研究生物学 圆形精子细胞，因为它们的使用会导致在形成单倍体细胞之前的表型。我们将 产生转基因小鼠系，其中所述转基因小鼠系的充分表征的圆形精子细胞特异性启动子 小鼠Acrv 1基因将驱动Cre重组酶的表达。该项目目标的实现将 提供了一种新的小鼠品系，使研究人员能够评估普遍表达的 圆形精子细胞分化（精子发生）过程中的基因。使用这种新的Cre-line（Acrv 1-Cre），我们 我建议测试这一假设，即普遍表达的TAR DNA/RNA结合蛋白43 千道尔顿（TDP-43）是精子发生所必需的。我们以前的工作已经确定TDP-43是 对男性生育能力至关重要TDP-43在圆形精子细胞中以及在步骤9-11的肩突中表达 精子细胞Acrv 1-Cre小鼠将允许我们测试其在精子发生过程中的作用。因此，创新 部分原因是，这项建议同时实现了两个目标。该提案符合所述目的， 融资机会PA-20-200。它将开发新的研究技术（圆形精子特异性Cre 删除小鼠品系），并作为试点和可行性研究，以关注TDP-43在精子发生中的作用。 这是一个小型研究项目，可以在资源有限的情况下在短时间内完成。

Project Summary The goal of this proposal is to generate and characterize a round spermatid-specific Cre deleter mouse line. Currently, there is no Cre-deleter mouse line available that targets deletion of a floxed gene specifically in round spermatids. The available testis germ cell Cre-lines delete floxed genes either in spermatogonia (Stra8- iCre) or in spermatocytes (Spo11-Cre, Hsp2a-Cre). These lines are not suitable for studying the biology of round spermatids because their use would cause a phenotype prior to the formation of haploid cells. We will generate a transgenic mouse line in which the well-characterized round spermatid-specific promoter of the mouse Acrv1 gene will drive the expression of Cre recombinase. Accomplishment of this project goal will provide a novel mouse line that will enable researchers to assess the function of ubiquitously expressed genes during round spermatid differentiation (spermiogenesis). Using this novel Cre-line (Acrv1-Cre), we propose to test the hypothesis that the ubiquitously expressed TAR DNA/RNA binding protein of 43 kilodaltons (TDP-43) is essential for spermiogenesis. Our previous work has established that TDP-43 is critical for male fertility. TDP-43 is expressed in the round spermatids as well as in the manchette of step 9-11 spermatids. The Acrv1-Cre mouse will allow us to test its role during spermiogenesis. Thus, the innovative part is that this proposal accomplishes two goals simultaneously. This proposal fulfils the stated purpose of the funding opportunity PA-20-200. It will develop new research technology (round spermatid-specific Cre deleter mouse line) and serve as a pilot and feasibility study to focus on the role of TDP-43 in spermiogenesis. It is a smallresearch project that can be carried out in a short period with limited resources.