The current technologies to place new DNA into specific locations in plant genomes are low frequency and error-prone, and this inefficiency hampers genome-editing approaches to develop improved crops^1,2. Often considered to be genome ‘parasites’, transposable elements (TEs) evolved to insert their DNA seamlessly into genomes^3,4,5. Eukaryotic TEs select their site of insertion based on preferences for chromatin contexts, which differ for each TE type^6,7,8,9. Here we developed a genome engineering tool that controls the TE insertion site and cargo delivered, taking advantage of the natural ability of the TE to precisely excise and insert into the genome. Inspired by CRISPR-associated transposases that target transposition in a programmable manner in bacteria^10,11,12, we fused the rice Pong transposase protein to the Cas9 or Cas12a programmable nucleases. We demonstrated sequence-specific targeted insertion (guided by the CRISPR gRNA) of enhancer elements, an open reading frame and a gene expression cassette into the genome of the model plant Arabidopsis. We then translated this system into soybean—a major global crop in need of targeted insertion technology. We have engineered a TE ‘parasite’ into a usable and accessible toolkit that enables the sequence-specific targeting of custom DNA into plant genomes.

目前将新 DNA 放入植物基因组特定位置的技术频率低且容易出错，这种低效率阻碍了开发改良作物的基因组编辑方法 ^1,2 。转座元件 (TE) 通常被认为是基因组“寄生虫”，它们经过进化可将其 DNA 无缝插入基因组 ^3,4,5 。真核 TE 根据对染色质环境的偏好来选择插入位点，每种 TE 类型 ^6,7,8,9 的染色质环境都不同。在这里，我们开发了一种基因组工程工具，利用 TE 精确切除和插入基因组的天然能力，控制 TE 插入位点和递送的货物。受到细菌 ^10,11,12 中以可编程方式定位转座的 CRISPR 相关转座酶的启发，我们将水稻 Pong 转座酶蛋白与 Cas9 或 Cas12a 可编程核酸酶融合。我们证明了增强子元件、开放阅读框和基因表达盒的序列特异性靶向插入（由 CRISPR gRNA 引导）到模式植物拟南芥的基因组中。然后，我们将该系统转化为大豆——一种需要定向插入技术的全球主要作物。我们将 TE“寄生虫”设计成一个可用且易于访问的工具包，能够将定制 DNA 序列特异性靶向植物基因组。