The role Y chromosome genes Prssly and Teyorf1 in male reproduction.

Y 染色体基因 Prssly 和 Teyorf1 在男性生殖中的作用。

• 批准号: 10337013
• 负责人: Monika A Ward
• 金额: $ 7.25万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-27 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10337013
• 关键词: Address; Automobile Driving; Back; Biological Assay; Blood-Testis Barrier; C-terminal; CRISPR/Cas technology; Chromosome Structures; Collaborations; Data; Defect; Development; Distal; Effectiveness; Electroporation; Evolution; FLAG peptide; Fertility; Fertilization; Future; Gene Proteins; Gene Targeting; Genes; Genetic; Genetic Transcription; Genome; Genotype; Germ Cells; Homologous Gene; Immunoblotting; Immunofluorescence Immunologic; In Vitro; Investigation; Knock-in; Knock-in Mouse; Knock-out; Knowledge; Link; Maintenance; Male Infertility; Mediating; Medical; Morphology; Mus; Oocytes; Open Reading Frames; Partner in relationship; Peptide Hydrolases; Phenotype; Physiological Processes; Play; Protein Family; Proteins; RNA Interference; Reporter; Reporting; Reproduction; Research Personnel; Resistance; Role; Serine; Serine Protease; Side; Sperm Motility; Spermatogenesis; System; Technology; Terminator Codon; Testing; Testis; Tissues; Transgenes; Translations; Y Chromosome; arm; cell motility; cell type; claudin 3; deletion analysis; gene function; interest; male; male fertility; medical specialties; member; molecular sequence database; mouse genome; positional cloning; protein expression; protein structure function; reproductive; reproductive function; sex; sex determination; sperm cell; sperm function; spermatogenic epithelium structure; sry Genes; structural genomics; transcription activator-like effector nucleases; ward; zygote; Address; Automobile Driving; Back; Biological Assay; Blood-Testis Barrier; C-terminal; CRISPR/Cas technology; Chromosome Structures; Collaborations; Data; Defect; Development; Distal; Effectiveness; Electroporation; Evolution; FLAG peptide; Fertility; Fertilization; Future; Gene Proteins; Gene Targeting; Genes; Genetic; Genetic Transcription; Genome; Genotype; Germ Cells; Homologous Gene; Immunoblotting; Immunofluorescence Immunologic; In Vitro; Investigation; Knock-in; Knock-in Mouse; Knock-out; Knowledge; Link; Maintenance; Male Infertility; Mediating; Medical; Morphology; Mus; Oocytes; Open Reading Frames; Partner in relationship; Peptide Hydrolases; Phenotype; Physiological Processes; Play; Protein Family; Proteins; RNA Interference; Reporter; Reporting; Reproduction; Research Personnel; Resistance; Role; Serine; Serine Protease; Side; Sperm Motility; Spermatogenesis; System; Technology; Terminator Codon; Testing; Testis; Tissues; Transgenes; Translations; Y Chromosome; arm; cell motility; cell type; claudin 3; deletion analysis; gene function; interest; male; male fertility; medical specialties; member; molecular sequence database; mouse genome; positional cloning; protein expression; protein structure function; reproductive; reproductive function; sex; sex determination; sperm cell; sperm function; spermatogenic epithelium structure; sry Genes; structural genomics; transcription activator-like effector nucleases; ward; zygote

Project Summary/Abstract The mammalian Y chromosome is a symbol of maleness as it encodes the Sry gene driving male sex determination, a battery of other genes thought to be involved in various aspects of male reproduction, and other genes playing roles of broadly expressed regulators of transcription, translation and protein stability. In spite of these clearly important functions, the knowledge linking the roles of specific Y chromosome genes to specific reproductive and other physiological processes remains limited. This is due to the unusual genomic structure of this chromosome, which for decades rendered it resistant to any targeting attempts. With the emergence of TALEN and CRISPR/Cas9 technologies addressing Y chromosome gene function directly became possible. The oddity of the Y chromosome structure made it also defiant to sequencing. The mouse Y chromosome sequence was finally defined in 2014, 12 years after the remainder of the mouse genome was characterized. When the mouse Y sequence was revealed two new single copy genes were identified on the Y short arm: Prssly (Y-linked serine-like protease) and Teyorf1 (testis expressed, chromosome Y open reading frame 1). These genes were classified as ‘acquired genes’ and were proposed to be the candidates for male fertility genes. Neither of these genes has been targeted and their function remains unknown. In Preliminary Data we show that Prssly and Teyorf1 expression is restricted to testicular postmeiotic germ cells. We also analyzed PRSSLY and TEYORF1 predicted protein structure and function and found that both predicted proteins resemble proteins that are known to play roles in spermatogenesis. PRSSLY belongs to the peptidase S1 family of proteins that include serine proteases PRSS21 and PRSS55 essential for sperm motility and sperm ability to fertilize oocytes. TEYORF1 belongs to the Claudin family of proteins members of which were reported important for assembly and maintenance of blood-testis barrier, a tissue barrier critical for normal spermatogenesis. Acquisition to the Y chromosome, testis specific expression, and resemblance to proteins of known reproductive functions prompted us to propose a hypothesis that Prssly and Teyorf1 play roles in male reproduction. We will address this hypothesis pursuing two specific aims: Aim 1: Generate Prssly and Teyorf1 knock-out (KO) and Prssly and Teyorf1 reporter tag knock-in (KI) mice using CRISPR/Cas9 system and zygote electroporation. Aim 2: Characterize phenotype of these mice focusing on assessment of fertility, sperm parameters, sperm function in vitro, and spermatogenesis progression. The combined results will allow to define whether Prssly and Teyorf1 play roles in spermatogenesis and fertility. The development of KI mice will open the door to future mechanistic investigations of these factors. The project will impact on understanding of Y chromosome gene function and genetic basis of male fertility. It will also contribute to discussions regarding the evolution of Y chromosomes and the importance of acquired Y genes.

项目摘要/摘要 哺乳动物y染色体是男性的象征，因为它编码了驱动男性确定的基因，一个 被认为与男性繁殖各个方面有关的其他基因的电池，其他基因扮演的角色 广泛表达的转录，翻译和蛋白质稳定性调节剂。尽管这些显然很重要 功能，将特定Y染色体基因与特定复制和其他生理学的作用联系起来的知识 过程仍然有限。这是由于该染色体的异常基因组结构，该结构数十年 它可以抵抗任何目标尝试。随着Talen和CRISPR/CAS9技术的出现 染色体基因功能直接成为可能。 Y染色体结构的奇数使其也反抗测序。小鼠y染色体序列是 最终定义在2014年，剩余的小鼠基因组的表征是在2014年的12年。当小鼠y序列 揭示了在Y短臂上鉴定了两个新的单拷贝基因：压力机（Y连锁丝氨酸蛋白酶）和 teyorf1（睾丸表示，染色体y开放阅读框1）。这些基因被归类为“获得的基因”，是 被认为是男性生育基因的候选者。这些基因均未针对且功能 仍然未知。 在初步数据中，我们表明，媒体和TEYORF1表达仅限于有证据的后生殖细胞。我们也是 分析的PRSLY和TEYORF1预测了蛋白质的结构和功能，发现两种蛋白质都类似于 已知在精子发生中起作用的蛋白质。 Prsly属于辣椒酶S1蛋白家族 包括串行蛋白酶PRSS21和PRSS55对于精子运动和精子施肥的能力必不可少。 teyorf1 属于蛋白质成员的克劳丁家族，这些家族对组装和维护很重要 血尾屏障，一种对正常精子发生至关重要的组织屏障。 获取Y染色体，睾丸特异性表达和与已知生殖功能的蛋白质相似 促使我们提出了一个假设，即媒体和Teyorf1在男性繁殖中扮演角色。我们将解决这个问题 假设追求两个具体目标： AIM 1：使用使用Pressly和Teyorf1淘汰赛（KO）以及Pressly和Teyorf1记者标签敲入（Ki）小鼠使用 CRISPR/CAS9系统和合子电穿孔。 目标2：表征这些小鼠的表型，专注于评估生育能力，精子参数，精子功能 体外和精子发生进展。 结合的结果将允许定义新闻和TEYORF1在精子发生和生育方面起作用。这 Ki小鼠的发展将为这些因素的未来机械调查打开大门。该项目将影响 了解Y染色体基因功能和男性生育能力的遗传基础。这也将有助于讨论 关注Y染色体的演变和获得的Y基因的重要性。