Collobert Mégane, Bocher Ozvan, Le Nabec Anaïs, Génin Emmanuelle, Férec Claude, Moisan Stéphanie
Univ. Brest, Inserm, EFS, UMR 1078, GGB, F-29200 Brest, France.
Department of Molecular Genetics and Reproduction Biology, CHRU Brest, F-29200 Brest, France.
Int J Mol Sci. 2021 Mar 5;22(5):2599. doi: 10.3390/ijms22052599.
About 8% of the human genome is covered with candidate -regulatory elements (cCREs). Disruptions of CREs, described as "-ruptions" have been identified as being involved in various genetic diseases. Thanks to the development of chromatin conformation study techniques, several long-range cystic fibrosis transmembrane conductance regulator () regulatory elements were identified, but the regulatory mechanisms of the gene have yet to be fully elucidated. The aim of this work is to improve our knowledge of the gene regulation, and to identity factors that could impact the gene expression, and potentially account for the variability of the clinical presentation of cystic fibrosis as well as -related disorders. Here, we apply the robust GWAS3D score to determine which of the introns could be involved in gene regulation. This approach highlights four particular introns of interest. Using reporter gene constructs in intestinal cells, we show that two new introns display strong cooperative effects in intestinal cells. Chromatin immunoprecipitation analyses further demonstrate fixation of transcription factors network. These results provide new insights into our understanding of the gene regulation and allow us to suggest a 3D locus structure in intestinal cells. A better understand of regulation mechanisms of the gene could elucidate cases of patients where the phenotype is not yet explained by the genotype. This would thus help in better diagnosis and therefore better management. These -acting regions may be a therapeutic challenge that could lead to the development of specific molecules capable of modulating gene expression in the future.
约8%的人类基因组覆盖有候选调控元件（cCREs）。被描述为“-ruptions”的CREs破坏已被确定与多种遗传疾病有关。由于染色质构象研究技术的发展，已鉴定出多个远距离囊性纤维化跨膜传导调节因子（）调控元件，但该基因的调控机制尚未完全阐明。这项工作的目的是增进我们对该基因调控的了解，确定可能影响该基因表达的因素，并有可能解释囊性纤维化以及相关疾病临床表现的变异性。在这里，我们应用强大的GWAS3D评分来确定该基因的哪些内含子可能参与基因调控。这种方法突出了四个特别感兴趣的内含子。在肠道细胞中使用报告基因构建体，我们表明两个新的内含子在肠道细胞中显示出强烈的协同作用。染色质免疫沉淀分析进一步证明了转录因子网络的固定。这些结果为我们理解该基因调控提供了新的见解，并使我们能够提出肠道细胞中的3D基因座结构。更好地理解该基因的调控机制可以阐明那些表型尚未由基因型解释的患者病例。因此，这将有助于更好的诊断，进而更好的治疗。这些顺式作用区域可能是一个治疗挑战，可能会导致未来能够调节基因表达的特定分子的开发。
