Department of Biology, Boston College, Chestnut Hill, Massachusetts 02467, USA; email:
Section on Eukaryotic Transposable Elements, Division of Molecular and Cellular Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA; email:
Annu Rev Genet. 2020 Nov 23;54:337-365. doi: 10.1146/annurev-genet-112618-043838. Epub 2020 Sep 4.
The goal of genomics and systems biology is to understand how complex systems of factors assemble into pathways and structures that combine to form living organisms. Great advances in understanding biological processes result from determining the function of individual genes, a process that has classically relied on characterizing single mutations. Advances in DNA sequencing has made available the complete set of genetic instructions for an astonishing and growing number of species. To understand the function of this ever-increasing number of genes, a high-throughput method was developed that in a single experiment can measure the function of genes across the genome of an organism. This occurred approximately 10 years ago, when high-throughput DNA sequencing was combined with advances in transposon-mediated mutagenesis in a method termed transposon insertion sequencing (TIS). In the subsequent years, TIS succeeded in addressing fundamental questions regarding the genes of bacteria, many of which have been shown to play central roles in bacterial infections that result in major human diseases. The field of TIS has matured and resulted in studies of hundreds of species that include significant innovations with a number of transposons. Here, we summarize a number of TIS experiments to provide an understanding of the method and explanation of approaches that are instructive when designing a study. Importantly, we emphasize critical aspects of a TIS experiment and highlight the extension and applicability of TIS into nonbacterial species such as yeast.
基因组学和系统生物学的目标是了解复杂的因素系统如何组装成途径和结构，这些途径和结构结合在一起形成生物体。从确定单个基因的功能中可以获得对生物过程的巨大理解进展，这一过程经典上依赖于对单个突变的特征描述。DNA 测序的进步使得越来越多的物种的全套遗传指令成为可能。为了理解这个数量不断增加的基因的功能，开发了一种高通量方法，该方法可以在单个实验中测量生物体基因组中基因的功能。大约 10 年前，当高通量 DNA 测序与转座子介导的诱变技术的进步相结合时，出现了一种称为转座子插入测序（TIS）的方法。在随后的几年中，TIS 成功地解决了有关细菌基因的基本问题，其中许多基因被证明在导致重大人类疾病的细菌感染中起着核心作用。TIS 领域已经成熟，并对包括许多转座子在内的数百个物种进行了研究。在这里，我们总结了一些 TIS 实验，以提供对该方法的理解，并解释在设计研究时具有指导意义的方法。重要的是，我们强调了 TIS 实验的关键方面，并强调了 TIS 向酵母等非细菌物种的扩展和适用性。
