Engineered geminivirus replicons enable rapid in planta directed evolution

IF 45.8 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Science Pub Date : 2025-10-02 DOI:10.1126/science.ady2167

Haocheng Zhu, Xu Qin, Leyan Wei, Dandan Jiang, Qiao Zhang, Wenqian Wang, Ronghong Liang, Rui Zhang, Kang Zhang, Guanwen Liu, Kevin Tianmeng Zhao, Kunling Chen, Jin-Long Qiu, Caixia Gao

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引用次数: 0

Abstract

Directed evolution can rapidly generate genetic variants with new and enhanced properties, yet efficient platforms for performing such evolution directly in plant cells have been lacking. We developed G eminivirus R eplicon- A ssisted in P lanta Directed E volution (GRAPE), a system that links gene function to geminivirus rolling circle replication (RCR) to enable high-throughput selection for desired activities. GRAPE supports the screening of up to 10 5 variants on a single Nicotiana benthamiana leaf within four days. Using GRAPE, we evolved the immune receptor NRC3 to resist inhibition by the nematode effector SPRYSEC15 and broadened the recognition spectrum of the rice immune receptor Pikm-1 to recognize all six alleles of the fungal effector AVR-Pik. GRAPE provides a rapid, scalable, and generalizable platform for directed evolution of diverse genes in plant cellular context.

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工程双病毒复制子使植物定向快速进化成为可能

定向进化可以迅速产生具有新的和增强特性的遗传变异，但在植物细胞中直接进行这种进化的有效平台一直缺乏。我们开发了G半病毒R eplicon- A，这是一种将基因功能与双病毒滚动环复制（RCR）联系起来的系统，可以实现高通量选择所需的活性。GRAPE支持在四天内筛选多达10个5个变异的单叶烟叶。利用葡萄，我们进化了免疫受体NRC3来抵抗线虫效应物SPRYSEC15的抑制，并拓宽了水稻免疫受体Pikm-1的识别谱，以识别真菌效应物AVR-Pik的所有6个等位基因。GRAPE为植物细胞环境下多种基因的定向进化提供了一个快速、可扩展和可推广的平台。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Science 综合性期刊-综合性期刊

CiteScore

61.10

自引率

0.90%

发文量

审稿时长

2.1 months

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