Exploring genotype-phenotype correlations in Sox10 mutations

项目描述：什么样的遗传变异会导致什么样的表型变化？SOX蛋白形成了一个转录因子家族，在胚胎发育和细胞动态平衡中具有关键功能，这些功能在脊椎动物中高度保守。由于其在多种细胞类型中的复杂作用，SOX突变的表型往往是不同的，即使是对于单一的细胞类型。虽然人们对它们的蛋白质结构和DNA结合特性知道得很多，但我们仍然不知道它们的结构和细胞功能之间的关系，即使在研究得很好的例子中，如Sox10(Pingault等人，2022，J.Med)，基因型-表型相关性仍然很模糊。吉内。59、105-114)。Sox10广泛表达于神经脊，这是一个高度多能性的祖细胞的胚胎种群，Sox10突变可能导致色素、听力、嗅觉和神经表型，单独或以各种组合的形式出现。表型的差异可能反映了突变(如功能的获得)或其他因素(如修饰基因座的存在)的微妙影响。我们已经证明了Sox10在斑马鱼和哺乳动物中的保守作用(例如Kelsh，2006，Bioessays 28,788-798)。此外，在同一团队监督的当前DTP学生的过程中，我们建立了一种产生CRISPR/Cas9诱导的精确基因组修改的方法，使用化学调制来加强同源定向修复(Zhang等人，2018，J.Biol)。化学。293,6611-6622；Aksoy等人，2019，Communications Biology，2,198)。这与斑马鱼胚胎的现成可获得性和表型特征(例如Alhasem等人，2022，eLife 11：e73550；Camargo-Sosa等人，2019，PLoS Genetics 15，e1007941)相结合，使斑马鱼成为在相对受限的遗传背景中探索特定突变变化的精确影响的理想系统。为了评估基因-表型差异，成功的申请者将从与疾病表型相关的人类变异中选择一系列广泛的斑马鱼x10等位基因；突变将被维持为杂合子，因为大多数突变可能是纯合子致命的。然后将对所有色素和神经细胞类型的显性和隐性表型进行定量表征。这将使我们能够解开这一重要发育调节因子目前鲜为人知但基本的结构-功能关系。这一跨学科项目将提供机会发展许多具体技能，包括斑马鱼遗传和养殖、包括原位杂交和免疫荧光在内的表型分析、共聚焦显微镜和光片显微镜、分子生物学和生物信息学。此外，学生还将受益于与巴黎想象研究所的老鼠和人类遗传学家的合作互动。

Project description: What genetic variants cause what phenotypic changes? The SOX proteins form a family of transcription factors with keyfunctions in embryonic development and cellular homeostasis, with these functions highly conserved across thevertebrates. Due to their complex roles in multiple cell-types, SOX mutant phenotypes are often diverse, even for a singlegene. Whilst much is known of their protein structure and DNA binding characteristics, we are still ignorant of therelationship between their structure and their cellular functions, with the genotype-phenotype correlation remainingobscure even in well-studied examples, such as SOX10 (Pingault et al., 2022, J. Med. Genet. 59, 105-114). SOX10 isexpressed widely in the neural crest, an embryonic population of highly multipotent progenitors, and SOX10 mutationsmay result in pigment, hearing, olfactory and neural phenotypes, individually or in various combinations. The variation inphenotypes may reflect subtle impacts of the mutations (e.g. gain of function), or other factors (e.g. presence of modifierloci). We have shown the conserved role for Sox10 in zebrafish and mammals (e.g. Kelsh, 2006, Bioessays 28, 788-798).Furthermore, in the course of a current DTP studentship supervised by same team, we have established a method forgenerating CRISPR/Cas9-induced precise genomic modifications, using chemical modulation to enhance Homology-Directed Repair (Zhang et al., 2018, J. Biol. Chem. 293, 6611-6622; Aksoy et al., 2019, Communications Biology, 2, 198).This, combined with the ready accessibility and phenotypic characterisation of zebrafish embryos (e.g. Alhashem et al.,2022, eLife 11:e73550; Camargo-Sosa et al., 2019, PLoS Genetics 15, e1007941), makes the zebrafish an ideal system toexplore the precise impacts of specific mutational changes in a relatively constrained genetic background.To assess the genotype-phenotype discrepancy, the successful applicant will create an extensive series of zebrafishsox10 alleles, selected from amongst the human variants linked to disease phenotypes; mutations will be maintained asheterozygotes, since most are likely to be homozygous lethal. Dominant and recessive phenotypes will then becharacterised quantitatively for all the pigment and neural cell-types. This will enable us to disentangle the currentlyobscure, but fundamental, structure-function relationships for this vital developmental regulatory factor. This interdisciplinary project will give an opportunity to develop numerous specific skillsets, including in zebrafish geneticsand husbandry, phenotypic analysis including by in situ hybridisation and immunofluorescence, confocal and lightsheet microscopy, molecular biology, and bioinformatics. In addition, the student will benefit from collaborativeinteraction with mouse and human geneticists at the Institut Imagine (Paris)