Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands.
Methods. 2012 Nov;58(3):204-11. doi: 10.1016/j.ymeth.2012.06.010. Epub 2012 Jun 19.
Chromosome conformation capture (3C) and 3C-based technology have revolutionized studies on chromosomal interactions and their role in gene regulation and chromosome organization. 3C allows the in vivo identification of physical interactions between chromosomal regions. Such interactions are shown to play a role in various aspects of gene regulation, for example transcriptional activation of genes by remote enhancer sequences, or the silencing by Polycomb-group complexes. The last few years the number of publications involving chromosomal interactions increased significantly. Until now, however, the vast majority of the studies reported are performed in yeast or animal systems. So far, studies on plant systems are extremely limited, possibly due to the plant-specific characteristics that hamper the implementation of the 3C technique. In this paper we provide a plant-specific 3C protocol, optimized for maize tissue, and an extensive discussion on (i) plant-specific adjustments to the protocol, and (ii) solutions to problems that may arise when optimizing the protocol for the tissue or plant of interest. Together, this paper should facilitate the application of 3C technology to plant tissue and stimulate studies on the 3D conformation of chromosomal regions and chromosomes in plants.
染色体构象捕获(3C)及其衍生技术极大地推动了对染色体相互作用及其在基因调控和染色体组织中的作用的研究。3C 允许在体内鉴定染色体区域之间的物理相互作用。这种相互作用被证明在基因调控的各个方面都起着重要作用,例如远程增强子序列对基因的转录激活,或 Polycomb 组复合物的沉默。近年来,涉及染色体相互作用的出版物数量显著增加。然而,到目前为止,绝大多数报道的研究都是在酵母或动物系统中进行的。到目前为止,对植物系统的研究极为有限,这可能是由于植物特有的特征阻碍了 3C 技术的实施。本文提供了一个针对玉米组织的植物特异性 3C 方案,并对(i)方案中的植物特异性调整,以及(ii)在针对感兴趣的组织或植物优化方案时可能出现的问题的解决方案进行了广泛的讨论。总的来说,本文应该有助于将 3C 技术应用于植物组织,并促进对植物中染色体区域和染色体的 3D 构象的研究。