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

生成新的 Cre-deleter 小鼠品系以研究精子发生

基本信息

批准号：
10668012
负责人：
PRABHAKARA P REDDI
金额：
$ 7.93万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10668012
关键词：
Acrosome Adult Affect Alleles Biogenesis Biology Breeding Chromatin Complementary DNA Couples DNA Ectopic Expression Enterobacteria phage P1 Cre recombinase Excision Feasibility Studies Funding Opportunities Gene Expression Gene Order Generations Genes Genotype Germ Cells Glycine Goals Haploid Cells Histopathology Human Pathology Incidence Infertility Knockout Mice LoxP-flanked allele Male Infertility Meiosis Meiotic Prophase I Modeling Mus Names Nuclear Nuclear Localization Signal Phenotype Physical condensation Pilot Projects Property Protamines RNA Recognition Motif RNA-Binding Proteins Reporter Reporter Genes Research Research Personnel Resources Reverse Transcriptase Polymerase Chain Reaction Role SPO11 gene Signaling Protein Specificity Sperm Motility Spermatids Spermatocytes Spermatogenesis Spermatogonia Spermiogenesis Tars Technology Testing Testis Tissues Transgenes Transgenic Mice Transgenic Organisms Work age group cell type conditional knockout improved innovation integration site male male fertility novel offspring prevent promoter protein TDP-43 red fluorescent protein reproductive spatiotemporal sperm cell sperm morphology transcription factor

项目摘要

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.

项目概要本提案的目标是生成并表征圆形精子细胞特异性 Cre 删除小鼠系。目前，还没有可用于特异性删除 floxed 基因的 Cre-deleter 小鼠品系。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 (Stra8- iCre) 或精母细胞 (Spo11-Cre、Hsp2a-Cre)。这些线不适合研究生物学圆形精子细胞，因为它们的使用会在单倍体细胞形成之前引起表型。我们将产生转基因小鼠品系，其中具有良好特征的圆形精子细胞特异性启动子小鼠 Acrv1 基因会驱动 Cre 重组酶的表达。本项目目标的实现将提供一种新的小鼠品系，使研究人员能够评估普遍表达的功能圆形精子细胞分化（精子发生）期间的基因。使用这个新颖的 Cre-line (Acrv1-Cre)，我们提议检验以下假设：43 中普遍表达的 TAR DNA/RNA 结合蛋白千道尔顿 (TDP-43) 对于精子发生至关重要。我们之前的工作已经确定 TDP-43 是对男性生育能力至关重要。 TDP-43 在圆形精子细胞以及步骤 9-11 的 manchette 中表达精子细胞。 Acrv1-Cre 小鼠将使我们能够测试其在精子发生过程中的作用。因此，创新的部分原因是该提案同时实现了两个目标。本提案实现了既定目的资助机会 PA-20-200。它将开发新的研究技术（圆形精子细胞特异性Cre 删除小鼠系）并作为试点和可行性研究，重点关注 TDP-43 在精子发生中的作用。这是一个小型研究项目，可以在短时间内利用有限的资源进行。