Genome editing beyond transgenes: DNA-free technologies in crop improvement
4 affiliations
- 1Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University
- 2Department of Agronomy, Directorate of Crop Management, Tamil Nadu Agricultural University
- 3Department of Vegetable Science, Horticultural College and Research Institute, Tamil Nadu Agricultural University
- 4Department of Plant Breeding, Swedish University of Agricultural Sciences
Abstract
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.
Related
- Beyond DNA—CRISPR technologies for epigenome editing and transcriptional regulation in disease: A scoping review
- Precision Editing of Cis-Regulatory Elements Drives Crops Quantitative Traits Genetic Improvement: From Functional Dissection to Intelligent Design.
- Functional improvement of dystrophic muscle by utrophin gene editing
- Short TALE-binding domains enable mitochondrial genome editing in Arabidopsis thaliana
- Prime editing for precise genome engineering and modulation of fungal metabolism
BibTeX
@article{senthilkumar2026genome,
title = {{Genome editing beyond transgenes: DNA-free technologies in crop improvement}},
author = {Roshini Senthilkumar and Rakshana Palaniswamy and Sakthi Ambothi Rathnasamy and Veera Ranjani Rajagopalan and Kokiladevi Eswaran and Radhamani Sengodan and Ashok Kumar Govindasamy and Raveendran Muthurajan and Selvaraju Kanagarajan and Sudha Manickam},
journal = {Frontiers in Plant Science},
year = {2026},
month = oct,
doi = {10.3389/fpls.2026.1942730},
url = {https://www.frontiersin.org/articles/10.3389/fpls.2026.1942730/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Genome editing beyond transgenes: DNA-free technologies in crop improvement
AU - Roshini Senthilkumar
AU - Rakshana Palaniswamy
AU - Sakthi Ambothi Rathnasamy
AU - Veera Ranjani Rajagopalan
AU - Kokiladevi Eswaran
AU - Radhamani Sengodan
AU - Ashok Kumar Govindasamy
AU - Raveendran Muthurajan
AU - Selvaraju Kanagarajan
AU - Sudha Manickam
PY - 2026
DA - 2026/10/05
JO - Frontiers in Plant Science
DO - 10.3389/fpls.2026.1942730
UR - https://www.frontiersin.org/articles/10.3389/fpls.2026.1942730/full
AB - 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.
SN - 1664-462X
ER -