Investigating sex differences in DDX3X mouse models

摘要 RNA结合蛋白DDX3X的从头突变导致DDX3X综合征。DDX3X 基因突变也与自闭症谱系障碍(ASD)有关，可能占到 女性不明原因的发育迟缓(DD)。然而，我们对DDX3X功能的理解 发育中的大脑是有限的。我们的资助旨在了解 DDX3X功能丧失和错义突变扰乱细胞功能。这份副刊将 关注与DDX3X相关的性二型表型。大多数从头突变发生在DDX3X 虽然观察到女性的病例数量在增加，但男性的病例数量有所增加。我们发现 女性体内DDX3X基因的完全缺失会导致严重的小头畸形。相比之下，完整的 男性的基因敲除，女性的单倍体不足。我们的数据表明，这些 雄性和雌性之间的差异可能是由于Paralog基因Ddx3y的表达发生了变化。 在这份补充材料中，我们将检验这一假设，即男性和女性的表型差异 与DDX3X相关的损失是由Paralog Ddx3y补偿的。为了测试这一点，我们将 使用单细胞转录组技术来量化细胞组成和相关的转录组变化 男性和女性胚胎大脑中的DDX3X缺失。此外，我们将使用击倒和 检测Ddx3y和DDX3X对胚胎脑功能冗余的抢救性实验 发展。这些实验将为理解DDX3X在 在细胞和分子水平上的神经发生。重要的是，这项提议旨在给 小鼠遗传学、胚胎学、测序、组织学和 显微镜。本副刊还包括一份全面的职业指导计划 发展。

Abstract De novo mutations in the RNA binding protein DDX3X result in DDX3X syndrome. DDX3X mutations are also associated with autism spectrum disorder (ASD) and may account for 1-3% of unexplained developmental delay (DD) in females. Yet, our understanding of DDX3X function in the developing brain is limited. Our funded grant aims to understand the mechanisms by which DDX3X loss of function and missense mutations perturb cellular function. This supplement will focus on sexually dimorphic phenotypes linked to DDX3X. Most de novo mutations are in DDX3X females though increasing numbers of cases in males have been observed. We have discovered that complete loss of Ddx3x in females causes striking microcephaly. In contrast, complete knockout in males phenocopies haploinsufficiency in females. Our data suggest that these differences between males and females may be due to altered expression of the paralog, Ddx3y. In this supplement, we will test the hypothesis that phenotypic differences in males and females associated with Ddx3x loss are due to compensation by the paralog, Ddx3y. To test this, we will use single cell transcriptomics to quantify cell composition and transcriptome changes associated with Ddx3x loss in males and female embryonic brains. Additionally, we will use knockdown and rescue experiments to test the functional redundancy of Ddx3y and Ddx3x for embryonic brain development. These experiments will give valuable new insights to understand roles for Ddx3x in neurogenesis at a cellular and molecular level. Importantly, this proposal is designed to give fundamental experimental training in mouse genetics, embryology, sequencing, histology and microscopy. This supplement also includes a comprehensive mentoring plan for career development.