Khokha Mustafa K, Loots Gabriela G
Department of Molecular and Cell Biology at the University of California, Berkeley, USA.
Brief Funct Genomic Proteomic. 2005 May;4(1):58-68. doi: 10.1093/bfgp/4.1.58.
Understanding the cis-regulatory architecture of metazoan organisms is the greatest challenge facing genome biology today. In vertebrate organisms, distinct sequence elements mediate transcriptional regulation and are scattered throughout the genome, either proximal or distal to promoters. The identification of transcriptional enhancers has proven rather difficult by conventional experimental approaches. In the past decade, the rapid generation of genomic sequences for multiple vertebrate organisms, accompanied by sophisticated comparative tools, has facilitated the identification of non-coding evolutionarily conserved regions that may encode cis-regulatory elements. Validating computational predictions and characterising cis-regulatory elements in vivo, however, has been a major bottleneck, mainly because the most commonly used organism for these experiments has been the mouse, and generating transgenic mice or modifying the mouse genome continues to be a labour-intensive, low-throughput, expensive process. This has led to the use of Xenopus, which holds great promise for high-throughput interrogation of putative cis-regulatory elements. In particular, Xenopus tropicalis may become particularly powerful for elucidating regulatory networks, chiefly because it is amenable to genetic manipulations, and its genome is being sequenced.
理解后生动物的顺式调控结构是当今基因组生物学面临的最大挑战。在脊椎动物中,不同的序列元件介导转录调控,它们分散在整个基因组中,位于启动子的近端或远端。通过传统实验方法鉴定转录增强子已被证明相当困难。在过去十年中,多种脊椎动物基因组序列的快速生成,伴随着复杂的比较工具,促进了对可能编码顺式调控元件的非编码进化保守区域的鉴定。然而,验证计算预测并在体内表征顺式调控元件一直是一个主要瓶颈,主要是因为这些实验最常用的生物体是小鼠,而生成转基因小鼠或修改小鼠基因组仍然是一个劳动密集型、低通量、昂贵的过程。这导致了非洲爪蟾的使用,它在高通量研究假定的顺式调控元件方面具有很大的前景。特别是热带爪蟾在阐明调控网络方面可能会变得特别强大,主要是因为它适合进行基因操作,并且其基因组正在测序。