Topological Organization of Drosophila Hox Genes Using DNA Fluorescent In Situ Hybridization
Topological Organization of Drosophila Hox Genes Using DNA Fluorescent In Situ Hybridization
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DOI：
10.1007/978-1-4939-1242-1_7
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Cavalli, Giacomo
中科院分区：
文献类型：
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作者：
Bantignies, Frederic;Cavalli, Giacomo
DNA荧光原位杂交（FISH）是在单细胞水平上研究基因组组织的首选方法。它最近被用来研究果蝇同源异型双胸复合体（BX-C）的拓扑结构，以及描述BX-C和果蝇足复合体（ANT-C）以及其他基因组位点之间的长程基因组相互作用。与免疫荧光相结合，FISH可用于研究同源异型基因与核亚区室的相对定位，如Polycomb组（PcG）体，转录工厂或核纤层。在这里，我们描述了两个3D-FISH协议，一个用于整个安装果蝇胚胎或幼虫光盘和果蝇培养的细胞。这两种方法都可以应用于任何单拷贝的目的基因座，并与免疫染色（FISH-1）兼容。
DNA fluorescent in situ hybridization (FISH) is the method of choice to study genomic organization at the single-cell level. It has been recently used to study the topological organization of the homeotic bithorax complex (BX-C) in Drosophila as well as to describe long-range genomic interactions between the BX-C and the Antennapedia complex (ANT-C), in addition to other genomic loci. Coupled with immunofluorescence, FISH can be used to study the relative positioning of homeotic genes with nuclear subcompartments, such as Polycomb-group (PcG) bodies, transcription factories, or the nuclear lamina. Here, we describe two multicolor 3D-FISH protocols; one for whole mount Drosophila embryos or larval discs and one for Drosophila-cultured cells. Both methods can be applied to any single copy locus of interest and are compatible with immunostaining (FISH-I).