CRISPR/Cas介导的无转基因编辑技术在植物中的应用研究进展

明美玲; 闫璐瑶; 易木林; 乔徐; 龙忠双; 付芳芳; 曹福亮

doi:10.12302/j.issn.1000-2006.202507034

明美玲, 闫璐瑶, 易木林, 乔徐, 龙忠双, 付芳芳, 曹福亮

南京林业大学学报（自然科学版） ›› 2026, Vol. 50 ›› Issue (4) : 27-42.

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南京林业大学学报（自然科学版） ›› 2026, Vol. 50 ›› Issue (4) : 27-42. DOI: 10.12302/j.issn.1000-2006.202507034

第二十八届中国科协年会———全球气候变化下的林草智能设计育种专题（执行主编　曹福亮　范国强　尹佟明　张怀清）
林学前沿

CRISPR/Cas介导的无转基因编辑技术在植物中的应用研究进展

明美玲 ¹ ,
闫璐瑶 ¹ ,
易木林 ¹ ,
乔徐 ² ,
龙忠双 ³ ,
付芳芳 ¹ ,
曹福亮 ¹^,^*

作者信息 +

Advances in CRISPR/Cas-mediated transgene-free genome editing in plants

Author information +

文章历史 +

摘要

转基因产品的生物安全监管问题始终是制约基因编辑植物市场化的核心因素,而发展无转基因编辑体系是推进农林业产业化的关键。尽管CRISPR/Cas系统及其衍生工具不断革新,其在植物中的递送效率与植株再生能力仍是创制基因编辑种质的关键瓶颈,严重限制该技术的普适性应用。系统综述CRISPR/Cas介导的基因编辑技术进展,涵盖不同类型基因编辑工具的开发应用、植物递送方法的创新突破以及无转基因编辑策略的最新进展。在此基础上,聚焦三大无转基因编辑技术路径:一是“不转入策略”,直接递送RNP、RNA复合体或RNA病毒载体,彻底避免外源DNA;二是“瞬时转化但不整合策略”,通过瞬时表达外源DNA或共编辑标记基因使编辑元件发挥作用后即降解,不整合至基因组;三是“稳定转化整合再清除策略”,通过基因删除技术、后代遗传分离或单倍体诱导等方式将整合的转基因成分去除。通过比较这些路径的原理与特性,为构建高效安全的植物无转基因编辑体系提供理论支撑和技术路线,以期突破基因编辑植物面临的产业化壁垒。

Abstract

The biosafety regulatory framework governing transgenic products has consistently emerged as a critical constraint impeding the commercialization trajectory of genome-edited plants. Consequently, the establishment of transgene-free editing technologies is of paramount importance for circumventing these regulatory impediments and facilitating the industrial application of plant genome editing. Despite the continuous and rapid innovation of CRISPR/Cas systems and their derivatives, the delivery efficiency and plant regeneration competence remain pivotal bottlenecks in the efficient generation of genome-edited germplasm, thereby severely constraining the broad applicability of these technologies. This review systematically delineates the advancements in CRISPR/Cas-mediated genome editing technologies, encompassing the development and practical application of diverse gene-editing toolkits, innovative breakthroughs in plant delivery methodologies, and the latest progresses in transgene-free editing strategies. Based on this comprehensive analysis, this review places specific emphasis on three principal transgene-free editing paradigms: (1) the “non-introduction” strategy, which entails the direct delivery of ribonucleoproteins (RNPs), RNA complexes, or RNA viral vectors to completely preclude the introduction of exogenous DNA; (2) the “transient transformation without integration” strategy, which achieves editing via the transient expression of exogenous DNA or co-editing of marker genes, ensuring that editing components execute their function and subsequently degrade without permanent genomic integration; (3) the “stable transformation, integration, and subsequent elimination” strategy, which employs gene excision technologies, progeny segregation, or haploid induction systems to eliminate integrated transgenic components. By systematically comparing the underlying principles and operational characteristics of these pathways, this review aims to provide robust theoretical underpinnings and feasible technical routes for establishing high-efficiency, high-safety transgene-free plant editing systems, with the ultimate objective of overcoming the industrialization barriers currently confronting genome-edited plants.

导出引用

明美玲, 闫璐瑶, 易木林, 等. CRISPR/Cas介导的无转基因编辑技术在植物中的应用研究进展[J]. 南京林业大学学报（自然科学版）. 2026, 50(4): 27-42 https://doi.org/10.12302/j.issn.1000-2006.202507034

Ming Meiling, Yan Luyao, Yi Mulin, et al. Advances in CRISPR/Cas-mediated transgene-free genome editing in plants[J]. Journal of Nanjing Forestry University (Natural Sciences Edition）. 2026, 50(4): 27-42 https://doi.org/10.12302/j.issn.1000-2006.202507034

中图分类号： S72;Q 943

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