Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes
Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes
复制标题
DOI：
10.1073/pnas.1213818110
复制
发表时间：
2012-12-26
影响因子：
11.1
通讯作者：
Wu, Chao-ting
中科院分区：
文献类型：
--
作者：
Beliveau, Brian J.;Joyce, Eric F.;Wu, Chao-ting
大量的观察结果表明，核结构和基因表达等过程之间的关系，导致了一些新的技术，询问染色体定位。其中一些技术重建了分子间的相互作用，而另一些技术则增强了我们原位可视化染色体的能力。在这里，我们描述了一种基于寡核苷酸和PCR的荧光原位杂交（FISH）策略和一个生物信息学平台，使这项技术能够扩展到任何基因组已被测序的生物体。寡核苷酸探针是可再生的、高效的，并且能够稳健地标记细胞培养物、固定组织和中期扩散中的染色体。我们的方法使研究人员能够精确控制他们的目标序列，并允许使用相同的基本协议对从数十个碱基到兆碱基的区域进行单次和双次成像。我们预计这项技术将导致增强的能力，可视化间期和中期染色体。
A host of observations demonstrating the relationship between nuclear architecture and processes such as gene expression have led to a number of new technologies for interrogating chromosome positioning. Whereas some of these technologies reconstruct intermolecular interactions, others have enhanced our ability to visualize chromosomes in situ. Here, we describe an oligonucleotide- and PCR-based strategy for fluorescence in situ hybridization (FISH) and a bioinformatic platform that enables this technology to be extended to any organism whose genome has been sequenced. The oligonucleotide probes are renewable, highly efficient, and able to robustly label chromosomes in cell culture, fixed tissues, and metaphase spreads. Our method gives researchers precise control over the sequences they target and allows for single and multicolor imaging of regions ranging from tens of kilobases to megabases with the same basic protocol. We anticipate this technology will lead to an enhanced ability to visualize interphase and metaphase chromosomes.