Construct design for precise DNA insertion in plants

Adnan Muzaffar; Md Torikul Islam; Adam M. Guss; Gerald A. Tuskan; Jin Gui Chen; Xiaohan Yang

doi:10.1016/j.tplants.2026.01.005

Construct design for precise DNA insertion in plants

Research output: Contribution to journal › Review article › peer-review

Abstract

Precise insertion of DNA sequences at targeted locations in plant genomes is pivotal for synthetic biology, genetics, and crop improvement. Construct design plays a critical role in achieving precise insertions, yet practical guidance remains limited. This review provides an in-depth overview of construct design principles and targeted DNA insertion (knock-in) strategies in plants. We assess the strengths, limitations, and construct requirements of current knock-in methods for specific applications, including short, large, and multifragment insertions. Additionally, we explore the potential of adopting advanced nonplant technologies to enhance knock-in efficiency and precision in plants. This review provides a valuable resource for facilitating the effective application of knock-in technologies to genetically improve crops with minimal off-target effects.

Original language	English
Journal	Trends in Plant Science
DOIs	https://doi.org/10.1016/j.tplants.2026.01.005
State	Accepted/In press - 2026

Funding

The writing of this manuscript was supported by the U.S. Department of Energy , Office of Science, Biological and Environmental Research Program, Genomic Science Program , as part of the Center for Bioenergy Innovation (CBI) under FWP ERKP886 , the Secure Ecosystem Engineering and Design (SEED) Scientific Focus Area under FWP ERKPA17 , and the Plant-Microbe Interfaces (PMI) Scientific Focus Area under FWP ERKP730 .

Keywords

DNA construct
genome editing
high-precision engineering
knock-in
plant biotechnology
synthetic biology

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10.1016/j.tplants.2026.01.005

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title = "Construct design for precise DNA insertion in plants",

abstract = "Precise insertion of DNA sequences at targeted locations in plant genomes is pivotal for synthetic biology, genetics, and crop improvement. Construct design plays a critical role in achieving precise insertions, yet practical guidance remains limited. This review provides an in-depth overview of construct design principles and targeted DNA insertion (knock-in) strategies in plants. We assess the strengths, limitations, and construct requirements of current knock-in methods for specific applications, including short, large, and multifragment insertions. Additionally, we explore the potential of adopting advanced nonplant technologies to enhance knock-in efficiency and precision in plants. This review provides a valuable resource for facilitating the effective application of knock-in technologies to genetically improve crops with minimal off-target effects.",

keywords = "DNA construct, genome editing, high-precision engineering, knock-in, plant biotechnology, synthetic biology",

author = "Adnan Muzaffar and Islam, \{Md Torikul\} and Guss, \{Adam M.\} and Tuskan, \{Gerald A.\} and Chen, \{Jin Gui\} and Xiaohan Yang",

note = "Publisher Copyright: {\textcopyright} 2026 The Author(s).",

year = "2026",

doi = "10.1016/j.tplants.2026.01.005",

language = "English",

journal = "Trends in Plant Science",

issn = "1360-1385",

publisher = "Cell Press",

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TY - JOUR

T1 - Construct design for precise DNA insertion in plants

AU - Muzaffar, Adnan

AU - Islam, Md Torikul

AU - Guss, Adam M.

AU - Tuskan, Gerald A.

AU - Chen, Jin Gui

AU - Yang, Xiaohan

PY - 2026

Y1 - 2026

N2 - Precise insertion of DNA sequences at targeted locations in plant genomes is pivotal for synthetic biology, genetics, and crop improvement. Construct design plays a critical role in achieving precise insertions, yet practical guidance remains limited. This review provides an in-depth overview of construct design principles and targeted DNA insertion (knock-in) strategies in plants. We assess the strengths, limitations, and construct requirements of current knock-in methods for specific applications, including short, large, and multifragment insertions. Additionally, we explore the potential of adopting advanced nonplant technologies to enhance knock-in efficiency and precision in plants. This review provides a valuable resource for facilitating the effective application of knock-in technologies to genetically improve crops with minimal off-target effects.

AB - Precise insertion of DNA sequences at targeted locations in plant genomes is pivotal for synthetic biology, genetics, and crop improvement. Construct design plays a critical role in achieving precise insertions, yet practical guidance remains limited. This review provides an in-depth overview of construct design principles and targeted DNA insertion (knock-in) strategies in plants. We assess the strengths, limitations, and construct requirements of current knock-in methods for specific applications, including short, large, and multifragment insertions. Additionally, we explore the potential of adopting advanced nonplant technologies to enhance knock-in efficiency and precision in plants. This review provides a valuable resource for facilitating the effective application of knock-in technologies to genetically improve crops with minimal off-target effects.

KW - DNA construct

KW - genome editing

KW - high-precision engineering

KW - knock-in

KW - plant biotechnology

KW - synthetic biology

UR - https://www.scopus.com/pages/publications/105032824609

U2 - 10.1016/j.tplants.2026.01.005

DO - 10.1016/j.tplants.2026.01.005

M3 - Review article

AN - SCOPUS:105032824609

SN - 1360-1385

JO - Trends in Plant Science

JF - Trends in Plant Science

ER -

Construct design for precise DNA insertion in plants

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