The Molecular Revolution in Cutaneous Biology: Identification of Skin Disease Genes
The Molecular Revolution in Cutaneous Biology: Identification of Skin Disease Genes
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DOI：
10.1016/j.jid.2016.11.019
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发表时间：
2017-05-01
影响因子：
6.5
通讯作者：
Choate, Keith A.
中科院分区：
文献类型：
--
作者：
Boyden, Lynn M.;Choate, Keith A.
在过去的30年里，发现与人类疾病相关的基因的能力显著扩大。从20世纪80年代的DNA标记和聚合酶链式反应的发展，90年代的荧光DNA标记和微阵列基因分型，到21世纪头10年人类全基因组的出版和高通量测序的兴起，每一次技术进步都加速了我们克隆和表征编码序列、将特征和疾病映射到特定的基因位点和发现致病突变的能力。在假说驱动的疾病基因功能鉴定中，特定的基因被作为假定的候选基因，基于现有的基因和疾病的生物学知识，有时借助模式生物研究。相比之下，位置克隆不需要疾病病理生物学或致病基因的先验知识。疾病基因定位克隆的时代始于20世纪80年代，当时一篇开创性的文章描述了如何将基因组DNA的变异用作遗传标记，以便在特定的染色体位置区分两种等位基因(Botstein等人，1980年)。这提供了通过家系追踪等位基因遗传的方法，从而寻找遗传模式与遗传病或特征相匹配的标记。这些标记可以是没有功能意义的多态，包括限制性片段长度多态、短串联重复序列(也称为微卫星)或单核苷酸多态。每种基因分型技术都在图谱效率和粒度方面提供了渐进式的进步。虽然标记本身通常不代表致病突变，但通过连锁分析检查它们的遗传模式可以为致病基因的染色体位置提供确凿的证据，其重要性可表示为对数赔率比分数。因此，随后发现致病突变的努力被限制在特定的基因座上，便于进行易于处理的搜索。在过去，一个成功的图谱项目往往需要克隆、测序和组装重叠的基因组DNA片段，以表征连锁区间内的基因，然后可以筛选突变。人类基因组计划的完成避免了这种费时费力的克隆和鉴定，并使从连锁分析快速过渡到突变发现成为可能。与此同时，随着测序技术的进步和成本的下降，突变检测从基于DNA迁移特性的方法(如异源双链和单链构象多态分析)转向通过Sanger或高通量DNA测序进行直接鉴定。
The past 30 years have witnessed a marked expansion in the power to discover genes relevant to human disease. From the development of DNA markers and PCR in the 1980s, fluorescent DNA labeling and microarray genotyping in the 1990s, and publication of the complete human genome and the rise of high-throughput sequencing in the 2000s, each technological advance has accelerated our ability to clone and characterize coding sequences, map traits and disorders to specific loci, and discover pathogenic mutations.In hypothesis-driven functional identification of disease genes, specific genes are examined as putative candidates on the basis of existing knowledge of the biology of the gene and disorder, sometimes aided by model organism studies. In contrast, positional cloning requires no prior knowledge about either disease pathobiology or the causative gene. The era of positional cloning of disease genes began in the 1980s with a groundbreaking article describing how variations in genomic DNA could be used as genetic markers that permit distinction of the two alleles at specific chromosomal loci (Botstein et al., 1980). This provides the means to track inheritance of alleles through pedigrees and thereby look for markers for which the inheritance pattern matches that of an inherited disease or trait. These markers can be polymorphisms without functional significance, including restriction fragment length polymorphisms, short tandem repeats (also known as microsatellites), or single nucleotide polymorphisms. Technologies to genotype each have provided incremental advances in mapping efficiency and granularity. Although the markers themselves do not usually represent pathogenic mutations, examination of their inheritance patterns via linkage analysis can provide definitive evidence of the chromosomal position of a causative gene, the significance of which may be represented as a logarithm-of-odds ratio score. Subsequent efforts to discover pathogenic mutations are thereby restricted to a specific locus, facilitating a tractable search. In the past, a successful mapping project often had to be followed by cloning, sequencing, and assembly of overlapping genomic DNA fragments to characterize the genes within the linkage interval, which could then be screened for mutations. The completion of the Human Genome Project obviated this laborious and time-consuming cloning and characterization and enabled rapid transition from linkage analysis to mutation discovery. Concurrently, as sequencing technologies advanced and costs declined, mutation detection moved from methods based on the migration properties of DNA (eg, heteroduplex and single-strand conformation polymorphism analysis) to direct identification via Sanger or high-throughput DNA sequencing.