Improving the specificity and throughput of automated analysis of chromatin fine structure in eukaryotic cells

提高真核细胞染色质精细结构自动分析的特异性和通量

• 批准号: BB/E005640/1
• 负责人: Constanze Bonifer
• 金额: $ 8.31万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE005640%2F1
• 关键词: Improving; specificity; throughput; automated; analysis

The molecular mechanism of gene expression is not fully understood, but the rules are the same for all organisms. One important discovery regards the role of chromatin. Chromatin is the name for the proteins compacting the one-meter long DNA molecule so that it fits into the cell nucleus. Active genes possess areas of reduced compaction. There are protein complexes pushing chromatin aside or modify it, so that genes can 'go to work'. The details of these processes are complicated and there are still many open questions. Therefore the chromatin structure of many genes needs to be studied so we can reach conclusions about general principles of the role of chromatin in gene expression, and how chromatin responds to signals coming from outside the cells. However, genes are very large and comprise many thousand and sometimes millions of base pairs and our current methods are too slow. We therefore urgently need methods that speed up the analysis of genes. We have developed a method that allows the rapid examination of the chromatin fine structure of entire gene loci, using a robot. However, although this has greatly improved our capacity to analyze genes, there is still a lot of room for improvement. This proposal aims at doing just that: we propose to further simplify existing technology and to make our assays more specific.

基因表达的分子机制尚不完全清楚，但所有生物体的规律都是相同的。一项重要的发现涉及到染色质的作用。染色质是一种蛋白质的名称，这种蛋白质将一米长的DNA分子压缩在一起，使其适合细胞核。活性基因具有压缩程度降低的区域。有一些蛋白质复合体将染色质推到一边或对其进行修饰，这样基因就可以“发挥作用”。这些过程的细节很复杂，仍有许多悬而未决的问题。因此，需要研究许多基因的染色质结构，从而得出染色质在基因表达中所起作用的一般原理，以及染色质如何对来自细胞外的信号做出反应。然而，基因非常大，由数千甚至数百万个碱基对组成，我们目前的方法太慢了。因此，我们迫切需要加快基因分析的方法。我们已经开发出一种方法，可以使用机器人快速检查整个基因座的染色质精细结构。然而，尽管这极大地提高了我们分析基因的能力，但仍有很大的改进空间。这项提议的目的正是为了做到这一点：我们提议进一步简化现有技术，并使我们的分析更加具体。

项目成果

PAP-LMPCR: an improved, sequence-selective method for the in vivo analysis of transcription factor occupancy and chromatin fine structure.

PAP-LMPCR：一种改进的序列选择性方法，用于转录因子占用和染色质精细结构的体内分析。

• DOI: 10.1007/978-1-60761-944-4_12
• 发表时间: 2011
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Ingram R

PAP-LMPCR for improved, allele-specific footprinting and automated chromatin fine structure analysis.

PAP-LMPCR 用于改进的等位基因特异性足迹和自动化染色质精细结构分析。

• DOI: 10.1093/nar/gkm1159
• 发表时间: 2008
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Ingram R