利用 CRISPR/Cas9 系统介导的高效烟草脆裂病毒诱导的番茄基因编辑。

Efficient tobacco rattle virus-induced gene editing in tomato mediated by the CRISPR/Cas9 system.

机构信息

College of Plant Protection, Yangzhou University, Yangzhou, China.

College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, China.

出版信息

Biotechnol J. 2024 May;19(5):e2400204. doi: 10.1002/biot.202400204.

DOI:10.1002/biot.202400204

PMID:38797722

Abstract

Plant virus-based sgRNA delivery strategy has been widely applied for efficient genome editing across various plant species, leveraging its significant advantages in the rapid expression and expansion of sgRNA through virus replication and movement. However, the efficacy of the virus-induced gene editing (VIGE) tool in tomato has yet to be explored. In this paper, we established a TRV-mediated CRISPR/Cas9 genome editing system in the somatic cells of tomato, reporting the validation of VIGE and evaluating the mutagenesis efficiency in both tomato leaves and fruits using high-throughput sequencing. The results demonstrated an approximate 65% efficiency of VIGE in tomato leaves for the selected target genes, with VIGE efficiency reaching up to 50% in tomato fruits. This research not only introduces an efficient tool for reverse genetics but also reveals substantial potential of VIGE in surpassing traditional tissue culture techniques for creating heritable mutations in tomato.

摘要

基于植物病毒的 sgRNA 递送策略已被广泛应用于各种植物物种的高效基因组编辑，利用其通过病毒复制和运动快速表达和扩展 sgRNA 的显著优势。然而，病毒诱导的基因编辑（VIGE）工具在番茄中的功效尚未得到探索。在本文中，我们在番茄体细胞中建立了一种基于 TRV 的 CRISPR/Cas9 基因组编辑系统，报告了 VIGE 的验证，并使用高通量测序评估了其在番茄叶片和果实中的诱变效率。结果表明，所选靶基因在番茄叶片中的 VIGE 效率约为 65%，在番茄果实中的 VIGE 效率高达 50%。这项研究不仅为反向遗传学引入了一种高效工具，还揭示了 VIGE 在超越传统组织培养技术方面的巨大潜力，可用于在番茄中创造可遗传的突变。