小柯机器人

中国科学院遗传与发育生物学研究所高彩霞等研究人员合作利用PrimeRoot编辑器精确整合植物基因组中的大型DNA序列。2023年4月24日，《自然—生物技术》杂志在线发表了这项成果。

研究人员报道了PrimeRoot，一种在植物中产生有针对性的精确大片段DNA插入的基因组编辑方法。第三代PrimeRoot编辑采用了优化的质粒编辑指导RNA设计，增强的植物质粒编辑和卓越的重组酶，以实现在植物基因组中精确的大型DNA插入，插入量可达11.1kb。研究人员展示了使用PrimeRoot在水稻中精确引入基因调控元件。

研究人员还将一个由PigmR组成的基因盒（该基因盒在Act1启动子的驱动下赋予稻瘟病抗性）整合到了北岳水稻的一个预测的基因组安全港位点，并获得了带有预期插入的编辑植物，效率为6.3%。研究人员发现，这些水稻植株具有更强的抗稻瘟性。这些结果确立了PrimeRoot是一种在植物中精确插入大段DNA的有望方法。

据悉，植物育种和合成生物学非常需要一种染色体插入大片段DNA的技术，以促进引入所需的农艺性状和信号及代谢途径。

附：英文原文

Title: Precise integration of large DNA sequences in plant genomes using PrimeRoot editors

Author: Sun, Chao, Lei, Yuan, Li, Boshu, Gao, Qiang, Li, Yunjia, Cao, Wen, Yang, Chao, Li, Hongchao, Wang, Zhiwei, Li, Yan, Wang, Yanpeng, Liu, Jun, Zhao, Kevin Tianmeng, Gao, Caixia

Issue&Volume: 2023-04-24

Abstract: A technique for chromosomal insertion of large DNA segments is much needed in plant breeding and synthetic biology to facilitate the introduction of desired agronomic traits and signaling and metabolic pathways. Here we describe PrimeRoot, a genome editing approach to generate targeted precise large DNA insertions in plants. Third-generation PrimeRoot editors employ optimized prime editing guide RNA designs, an enhanced plant prime editor and superior recombinases to enable precise large DNA insertions of up to 11.1kilobases into plant genomes. We demonstrate the use of PrimeRoot to accurately introduce gene regulatory elements in rice. In this study, we also integrated a gene cassette comprising PigmR, which confers rice blast resistance driven by an Act1 promoter, into a predicted genomic safe harbor site of Kitaake rice and obtain edited plants harboring the expected insertion with an efficiency of 6.3%. We found that these rice plants have increased blast resistance. These results establish PrimeRoot as a promising approach to precisely insert large segments of DNA in plants.

DOI: 10.1038/s41587-023-01769-w

Source: https://www.nature.com/articles/s41587-023-01769-w

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex