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

Ming Meiling, Yan Luyao, Yi Mulin, Qiao Xu, Long Zhongshuang, Fu Fangfang, Cao Fuliang

Journal of Nanjing Forestry University (Natural Sciences Edition） ›› 2026, Vol. 50 ›› Issue (4) : 27-42.

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

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

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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.

Key words

CRISPR/Cas / genome editing tools / delivery methods / transgene-free / biosafety

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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

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