Abstract:
Transposons are mobile genetic elements evolved to execute highly efficient integration of their genes into the genomes of their host cells. These natural DNA transfer vehicles have been harnessed as experimental tools for stably introducing a wide variety of foreign DNA sequences, including selectable marker genes, reporters, shRNA expression cassettes, mutagenic gene trap cassettes, and therapeutic gene constructs into the genomes of target cells in a regulated and highly efficient manner. Given that transposon components are typically supplied as naked nucleic acids (DNA and RNA) or recombinant protein, their use is simple, safe, and economically competitive. Thus, transposons enable several avenues for genome manipulations in vertebrates, including transgenesis for the generation of transgenic cells in tissue culture comprising the generation of pluripotent stem cells, the production of germline-transgenic animals for basic and applied research, forward genetic screens for functional gene annotation in model species and therapy of genetic disorders in humans. This review describes the molecular mechanisms involved in transposition reactions of the three most widely used transposon systems currently available (Sleeping Beauty, piggyBac, and Tol2), and discusses the various parameters and considerations pertinent to their experimental use, highlighting the state-of-the-art in transposon technology in diverse genetic applications.
摘要:
转座子是进化的可移动遗传元件,可将其基因高效整合到其宿主细胞的基因组中。这些天然DNA转移载体已被用作稳定引入各种外源DNA序列的实验工具。包括选择标记基因,记者,shRNA表达盒,诱变基因捕获盒,和治疗基因构建到靶细胞的基因组以受调节和高效的方式。鉴于转座子成分通常作为裸核酸(DNA和RNA)或重组蛋白提供,它们的使用很简单,安全,经济竞争力。因此,转座子为脊椎动物的基因组操作提供了几种途径,包括用于在组织培养中产生转基因细胞的转基因,包括多能干细胞的产生,用于基础和应用研究的种系转基因动物的生产,用于模型物种功能基因注释的正向遗传筛选和人类遗传疾病的治疗。这篇综述描述了目前可用的三种最广泛使用的转座子系统(睡美人,piggyBac,andTol2),并讨论了与实验使用相关的各种参数和注意事项,强调转座子技术在不同遗传应用中的最新水平。