Genetic tools and mutagenesis strategies based on transposable elements are currently under development with a vision to link primary DNA sequence information to gene functions in vertebrate models. By virtue of their inherent capacity to insert into DNA, transposons can be developed into powerful tools for chromosomal manipulations. Transposon-based forward mutagenesis screens have numerous advantages including high throughput, easy identification of mutated alleles, and providing insight into genetic networks and pathways based on phenotypes. For example, the Sleeping Beauty transposon has become highly instrumental to induce tumors in experimental animals in a tissue-specific manner with the aim of uncovering the genetic basis of diverse cancers. Here, we describe a battery of mutagenic cassettes that can be applied in conjunction with transposon vectors to mutagenize genes, and highlight versatile experimental strategies for the generation of engineered chromosomes for loss-of-function as well as gain-of-function mutagenesis for functional gene annotation in vertebrate models, including zebrafish, mice, and rats.

目前正在开发基于转座因子的遗传工具和诱变策略，以期将主要的DNA序列信息与脊椎动物模型中的基因功能联系起来。凭借其固有的插入DNA的能力，转座子可以发展成为用于染色体操作的强大工具。基于转座子的正向诱变筛选具有许多优势，包括高通量，易于识别突变的等位基因，并提供对基于表型的遗传网络和途径的洞察力。例如《睡美人》转座子已成为在组织中特异性诱导实验动物肿瘤的重要工具，目的是揭示各种癌症的遗传基础。在这里，我们描述了一系列诱变盒，可以将其与转座子载体结合使用来诱变基因，并着重介绍针对功能丧失和功能获得诱变的工程化染色体生成的通用实验策略脊椎动物模型（包括斑马鱼，小鼠和大鼠）中的基因注释。