Construct design for precise DNA insertion in plants

Trends Plant Sci. 2026 Jul;31(7):887-910. doi: 10.1016/j.tplants.2026.01.005. Epub 2026 Mar 17.

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.

Publication types

  • Review
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Crops, Agricultural / genetics
  • DNA, Plant* / genetics
  • Gene Knock-In Techniques* / methods
  • Genetic Engineering* / methods
  • Genome, Plant / genetics
  • Mutagenesis, Insertional* / methods
  • Plants* / genetics
  • Plants, Genetically Modified / genetics

Substances

  • DNA, Plant