Exploiting natural variants in potassium channel genes to understand their roles in neural function, behaviour and development

利用钾通道基因的自然变异来了解它们在神经功能、行为和发育中的作用

• 批准号: 2893712
• 金额: --
• 依托单位: University of Portsmouth
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2893712
• 关键词: Exploiting; natural; variants; potassium; channel

Rationale and Impact: There are around 100 genes encoding potassium channels, and around 40% of these are associated with diseases, generally displaying developmental delay, ataxia and epilepsy.We aim to systematically assess the function of the remaining potassium channel genes to better understand this gene family. This understanding will have impact, the data will help improve diagnosis and interventions for people who have pathogenic variants in these genes.We recently identified a novel syndrome characterised by ataxia and developmental delay caused by variants in a potassium channel gene, KCNC4. Our innovative approach combined bioinformatics investigation of human variation, molecular modelling of channel function, and recapitulation of the human phenotype in the Xenopus model organism to link the gene to the disorder and understand its molecular basis. We will extend this successful approach to other potassium channels.Approach:Bioinformatics: Whole genome sequences from patients with rare disorders in the 100,000 Genomes Project (100KGP) and NHS Genomic Medicine Service will be interrogated for damaging variants in potassium channel genes. These will be prioritised based on bioinformatics tools, number of probands and their phenotypes to determine the most likely disease causing genes.Ion channel function: Using well established electrophysiological assays, the impact of the detected variants on ion channel function will be measured relative to the wild-type channel using patch clamp in HEK cells. This will establish whether the variants impact channel function, and what the nature of that disruption is.Xenopus models: Promising genes will be knocked-out using CRISPR in Xenopus, and the resultant morphological, physiological and behavioural changes systematically assessed. This will determine the normal function of the gene in the whole organism.Candidate qualities: An ability to engage with a multi-disciplinary project within an existing, successful collaboration. Specifically, an interest in bioinformatics, electrophysiology, molecular genetics and phenotyping.

原理和影响：大约有 100 个编码钾通道的基因，其中大约 40% 与疾病相关，通常表现为发育迟缓、共济失调和癫痫。我们的目标是系统评估其余钾通道基因的功能，以更好地了解这一点基因家族。这种理解将会产生影响，这些数据将有助于改善对这些基因中存在致病性变异的人的诊断和干预。我们最近发现了一种新的综合征，其特征是由钾通道基因 KCNC4 变异引起的共济失调和发育迟缓。我们的创新方法结合了人类变异的生物信息学研究、通道功能的分子建模以及非洲爪蟾模型生物体中人类表型的重演，以将基因与疾病联系起来并了解其分子基础。我们将把这种成功的方法推广到其他钾离子通道。方法：生物信息学：来自 100,000 基因组计划 (100KGP) 和 NHS 基因组医学服务中罕见疾病患者的全基因组序列将被询问钾离子通道基因的破坏性变异。将根据生物信息学工具、先证者数量及其表型对这些进行优先排序，以确定最可能的致病基因。 离子通道功能：使用完善的电生理学测定，将测量检测到的变异对离子通道功能的影响相对于在 HEK 细胞中使用膜片钳的野生型通道。这将确定这些变异是否影响通道功能，以及这种破坏的本质是什么。 非洲爪蟾模型：在非洲爪蟾中使用 CRISPR 敲除有前途的基因，并系统地评估由此产生的形态、生理和行为变化。这将决定该基因在整个生物体中的正常功能。候选人素质：在现有的成功合作中参与多学科项目的能力。具体来说，对生物信息学、电生理学、分子遗传学和表型分析感兴趣。