Frontiers in plant scienceOct 20, 2026

DNA-free genome editing methods for improving crops

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Oct 21, 2026

Abstract

DNA-free genome editing using CRISPR/Cas9 offers a precise method for crop improvement without stable transgene integration.

This approach enables targeted genetic modifications that do not leave foreign genes in the plant's genome.
After editing, the introduced elements are quickly degraded, resulting in plants that are indistinguishable from those produced by traditional breeding.
Various delivery systems for the CRISPR/Cas9 components have been developed for major crop species, including protoplast transfection and nanoparticle methods.
Enhancements in traits such as disease resistance, nutritional quality, and stress tolerance have been achieved in multiple crop types.
Recent advances like base editing and multiplex gene editing may improve the precision and effectiveness of this technique.
Wider application of this method could depend on advancements in plant regeneration and regulatory frameworks.

Simplified

Conventional and transgenic approaches to crop improvement are increasingly constrained by biosafety concerns, stringent regulatory frameworks, and limited public acceptance, necessitating new precision breeding methods. DNA-free genome editing using the CRISPR/Cas9 approach has emerged as a transformative approach that enables precise, targeted genetic modifications without the stable integration of a foreign gene (transgene). This strategy transiently introduces CRISPR/Cas9 ribonucleoprotein (RNP) complexes, or in vitro transcribed mRNA, into plant cells. This review focuses primarily on CRISPR/Cas-mediated DNA-free genome editing and its applications in crop improvement. After targeted genome cleavage, the editing elements are rapidly degraded, leaving only the desired genomic modification and resulting in plants indistinguishable from those developed through conventional breeding. To achieve targeted, DNA-free genome editing, researchers have developed numerous molecular platforms and delivery systems, such as protoplast transfection, biolistic bombardment, nanoparticle-based systems, and emerging pollen- and meristem-targeted systems, for major crop species. Researchers have successfully enhanced several notable traits, including disease and pest resistance, nutritional quality, abiotic stress tolerance, and agronomic performance, in cereals, vegetables, oilseeds, and perennial crops. Recent advancements, including base editing, prime editing, and multiplex gene editing, are further expanding the precision, efficiency, and scope of DNA-free genome editing. Overall, CRISPR/Cas-mediated DNA-free genome editing offers a route to crop improvement that avoids stable transgene integration, though its wider deployment depends on progress in regeneration, molecular validation, and regulatory convergence.

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DNA-free genome editing methods for improving crops

Abstract

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