The evolving landscape of precise DNA insertion in plants

Tien Van Vu (), Ngan Thi Nguyen, Jihae Kim, Yeon Woo Sung, Woo Sik Chung and Jae-Yean Kim ()
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Tien Van Vu: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center
Ngan Thi Nguyen: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center
Jihae Kim: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center
Yeon Woo Sung: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center
Woo Sik Chung: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center
Jae-Yean Kim: Gyeongsang National University, Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center

Nature Communications, 2025, vol. 16, issue 1, 1-17

Abstract: Abstract Precise DNA insertion into plant genomes is central to advancing crop improvement and synthetic biology. CRISPR-Cas systems have enabled programmable DNA integration using tools such as gene targeting (GT), prime editing (PE), and recombinase- or transposase-based platforms. These tools are transitioned from theoretical concepts to practical applications, supporting applications like in-locus protein tagging, regulatory element engineering, and multi-gene stacking. Key challenges persist, such as inefficient large-fragment insertion, delivery barriers, and regulatory hurdles. This review traces the evolution from random to CRISPR-Cas-based systems, analyzes current limitations, and discusses emerging solutions paving the way for predictable DNA insertion in modern plant biotechnology.

Date: 2025
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DOI: 10.1038/s41467-025-66715-7

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