Roles of X- and Y-palindromic Genes in Mammalian Fertility

X 和 Y 回文基因在哺乳动物生育力中的作用

• 批准号: 9906246
• 负责人: Jacob L Mueller
• 金额: $ 33.64万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906246
• 关键词: Address; Antibodies; Architecture; Biological; Biology; Characteristics; Chromosomes; Collection; Complex; DNA; DNA Sequence; Defect; Development; Engineering; Excision; Exhibits; Family member; Fertility; Fertilization; Funding; Gene Family; Genes; Genetic; Genome engineering; Genomics; Germ Cells; Human; Immunoprecipitation; Individual; Infertility; Knock-out; Knockout Mice; Laboratories; Link; Male Infertility; Mediating; Meiosis; Methodology; Molecular; Mus; Mutation; Nature; Proteins; RNA; Reproducibility; Research; Role; Sex Chromosomes; Sex Ratio; Sperm Count Procedure; Spermatids; Spermatogenesis; Spermatogenic Cell; System; Testing; Testis; X Chromosome; Y Chromosome; base; experimental study; gene function; genetic manipulation; human male; improved; in vivo; innovation; insight; knock-down; loss of function; male; male fertility; mammalian genome; member; novel; screening; sperm cell; structural genomics

Project Summary Mammalian sex chromosomes are enriched with large palindromic sequences harboring gene families expressed almost exclusively in spermatogenic cells, however the molecular functions of these palindromic genes remain largely unknown. Understanding the mechanistic functions of individual palindromic gene families will provide novel insights into genetic factors that contribute to spermatogenic defects (abnormalities in the development of testicular germ cells). The objective of this proposal is to precisely delete individual X- and Y-palindrome gene families to determine their molecular functions during spermatogenesis. This proposal addresses whether deletion of all copy members of an individual X- or Y-palindromic gene family results in spermatogenic defects. This proposition is based on the observation that deletions that remove all members of X- or Y-palindromic gene families result in spermatogenic defects. However, each of these previously studied deletions remove more genes than just an individual palindromic gene family, making it difficult to determine the contribution of individual palindromic gene families to spermatogenic defects. Therefore, we propose to generate precise deletions that remove all members of individual X- or Y-palindromic gene families in mice to understand their role in spermatogenesis. As proof of feasibility, we have generated two, independent, megabase-sized deletions of the Slx and Slxl1 X-palindromic gene families and show that male mice carrying deletions of both gene families are infertile, exhibiting a specific post-meiotic spermatogenic defect (Preliminary Studies). This proposal addresses the importance of X- and Y-palindromic genes via two specific aims: 1) Determine the role of the Slx and Slxl1 X-palindromic gene families in post-meiotic spermatogenesis in male mice carrying deletions of both the Slx and Slxl1 palindrome arrays; 2) Determine the contribution of four additional X- and Y-palindromic gene families in spermatogenesis by individually deleting each of the four distinct gene families. To genetically dissect the functions of individual X- and Y-palindromic gene families in mice, we have developed new chromosome engineering methodologies to efficiently delete large palindromic regions in vivo. Each of the X- and Y-palindromic gene families in mice will be systematically characterized for defects in spermatogenesis. The functions of individual X- and Y-palindromic gene families have been poorly studied in humans and mice because of their recent discovery and complex genomic architecture. The proposed experiments will provide an improved understanding of the roles of individual X- and Y-palindromic genes in male fertility. The fact that X- and Y-palindromes harbor testis-specific gene families makes them ideal candidates for uncovering new genetic factors responsible for human spermatogenic defects. By understanding the molecular functions of X- and Y-palindromic gene families, important insights can be gained into their role in enabling spermatogenesis and how mutations in X- and Y-palindromic gene families disrupt spermatogenesis.

项目摘要 哺乳动物的性染色体上富含大量的回文序列， 几乎只在生精细胞中表达，然而这些回文的分子功能 基因在很大程度上仍是未知的。理解单个回文基因的机制功能 家庭将提供新的见解遗传因素，有助于精子发生缺陷（异常 睾丸生殖细胞的发育）。这一建议的目的是准确地删除个别X- 和Y回文基因家族，以确定其在精子发生过程中的分子功能。这项建议 解决了单个X-或Y-回文基因家族的所有拷贝成员的缺失是否导致 生精缺陷这个命题是基于这样的观察，删除所有成员的删除， X或Y回文基因家族导致精子发生缺陷。然而，之前研究的每一种 缺失删除的基因比单个回文基因家族更多，这使得很难确定 单个回文基因家族对生精缺陷的贡献。因此，我们建议 产生精确的删除，删除小鼠中单个X或Y回文基因家族的所有成员， 了解它们在精子发生中的作用。作为可行性的证明，我们已经生成了两个独立的， Slx和Slxl 1 X-回文基因家族的兆碱基大小的缺失，并显示携带 两个基因家族的缺失都是不育的，表现出特异性的减数分裂后精子发生缺陷（初步研究）。 研究）。该建议通过两个具体目标来解决X和Y回文基因的重要性：1） 确定Slx和Slxl 1 X回文基因家族在男性减数分裂后精子发生中的作用 携带Slx和Slxl 1回文序列缺失的小鼠; 2）确定四种基因的贡献 另外的X-和Y-回文基因家族在精子发生中通过单独删除四个 不同的基因家族为了从遗传学上剖析单个X和Y回文基因家族在基因组中的功能， 小鼠，我们已经开发了新的染色体工程方法，以有效地删除大回文 体内区域。小鼠中的每个X-和Y-回文基因家族将被系统地表征， 精子发生缺陷。单个X和Y回文基因家族的功能一直很差， 因为它们最近的发现和复杂的基因组结构，我们在人类和小鼠中进行了研究。的 提出的实验将提供一个更好的理解个人的X和Y回文的作用， 男性生育力的基因事实上，X-和Y-回文酶具有睾丸特异性基因家族， 是发现导致人类生精缺陷的新遗传因子的理想候选者。通过 了解X和Y回文基因家族的分子功能，可以获得重要的见解 研究它们在精子发生中的作用，以及X和Y回文基因家族的突变如何破坏 精子发生