New fluorescent probes for labelling nucleic acids

用于标记核酸的新型荧光探针

• 批准号: BB/L01811X/1
• 负责人: Tom Brown
• 金额: $ 18.99万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL01811X%2F1
• 关键词: New; fluorescent; probes; labelling; nucleic

The human body is composed of billions of cells of different types. Each cell type (e.g. skin cells, liver cells, red blood cells) has its own specific roles. Many diseases arise from a breakdown or defect in the complex biochemistry within specific types of cells. A clinically important example of this is thalassaemia. The alpha and beta thalassaemias are common severe inherited forms of anaemia caused by imbalance of alpha and beta globin gene expression which affects the production of haemoglobin, the protein that carries oxygen through the body. Current therapy involves lifelong blood transfusion and iron chelation therapy, but these are invasive and only moderately effective. Consequently new therapies to correct globin chain imbalance are urgently needed. A crucial requirement in the development of new treatments is a detailed understanding of the regulation of alpha and beta globin gene expression, i.e. why these genes do not make their products in the correct amounts in patients suffering from thalassaemia. It is now recognized that the ability of many genes to make proteins is regulated by enhancer elements in their DNA. These are binding sites for proteins that switch genes on or off, and mutations at these sites underlie many human diseases. It has been shown by special fluorescence cell imaging techniques that enhancers and promoters can be located far apart on DNA, but come together when they are controlling protein synthesis. However it is not clear how this physical interaction exerts its effect. To analyse this we must obtain much clearer physical pictures of the process to determine how dynamic this enhancer-promoter association is, when it occurs and how long it persists. This will require new methods for high-resolution imaging of genomic DNA using small and intensely bright pieces of DNA (fluorescent probes). These probes will be much brighter than those currently available. They will be synthesised chemically by high-throughput automated methods using a modular approach which will greatly reduce the cost of their manufacture. This new approach to DNA and RNA imaging is generic and will be used in a wide variety of clinical and research applications by scientists working in academia, research institutes and the pharmaceutical industry.

人体由数十亿个不同类型的细胞组成。每种细胞类型(如皮肤细胞、肝细胞、红细胞)都有其特定的作用。许多疾病是由特定类型细胞内复杂生物化学的故障或缺陷引起的。临床上的一个重要例子就是地中海贫血症。阿尔法和贝塔地中海贫血是一种常见的严重遗传性贫血，由阿尔法和贝塔珠蛋白基因表达失衡引起，这会影响血红蛋白的产生，血红蛋白是一种在体内输送氧气的蛋白质。目前的治疗方法包括终生输血和铁络合疗法，但这些都是侵入性的，而且只有适度的效果。因此，迫切需要新的治疗方法来纠正珠蛋白链失衡。开发新的治疗方法的一个关键要求是详细了解α和β珠蛋白基因表达的调节，即为什么这些基因在地中海贫血症患者中不能产生正确数量的产物。现在人们认识到，许多基因制造蛋白质的能力是由其DNA中的增强子元件调节的。这些是开启或关闭基因的蛋白质的结合位点，这些位点的突变是许多人类疾病的基础。特殊的荧光细胞成像技术表明，增强子和启动子可以位于DNA上很远的地方，但当它们控制蛋白质合成时，它们会聚集在一起。然而，目前还不清楚这种物理相互作用是如何发挥作用的。为了分析这一点，我们必须对这一过程获得更清晰的物理图像，以确定这种增强子-启动子关联的动态程度、何时发生以及持续多久。这将需要使用小而明亮的DNA片段(荧光探针)对基因组DNA进行高分辨率成像的新方法。这些探测器将比目前可用的探测器明亮得多。它们将通过使用模块化方法的高通量自动化方法进行化学合成，这将大大降低它们的制造成本。这种DNA和RNA成像的新方法是通用的，将被学术界、研究机构和制药行业的科学家用于广泛的临床和研究应用。

项目成果

A tissue-specific self-interacting chromatin domain forms independently of enhancer-promoter interactions.

• DOI: 10.1038/s41467-018-06248-4
• 发表时间: 2018-09-21
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Brown JM;Roberts NA;Graham B;Waithe D;Lagerholm C;Telenius JM;De Ornellas S;Oudelaar AM;Scott C;Szczerbal I;Babbs C;Kassouf MT;Hughes JR;Higgs DR;Buckle VJ
• 通讯作者: Buckle VJ

Chromatin arranges in chains of mesoscale domains with nanoscale functional topography independent of cohesin.

• DOI: 10.1126/sciadv.aba8811
• 发表时间: 2020-09
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Miron E;Oldenkamp R;Brown JM;Pinto DMS;Xu CS;Faria AR;Shaban HA;Rhodes JDP;Innocent C;de Ornellas S;Hess HF;Buckle V;Schermelleh L
• 通讯作者: Schermelleh L

Chromatin arranges in chains of mesoscale domains with nanoscale functional topography independent of cohesin

染色质排列成具有纳米级功能拓扑的中尺度结构域链，与粘连蛋白无关

• DOI: 10.1101/566638
• 发表时间: 2019
• 作者: Miron E