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Updated: Dec 15, 2025

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Targeted, efficient sequence insertion and replacement in rice.

Yuming Lu1, Yifu Tian1, Rundong Shen1

  • 1Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China.

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|July 8, 2020
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Summary
This summary is machine-generated.

Researchers developed new CRISPR-Cas9 methods for inserting long DNA sequences and genes into rice, advancing crop improvement and functional genomics research.

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Area of Science:

  • Plant Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • CRISPR-Cas9 technology enables various genome modifications in plants, including insertions, deletions, and base changes.
  • Existing methods for inserting long sequences or genes in plants are limited and often require selection markers, hindering functional genomics and crop trait improvement.

Purpose of the Study:

  • To develop versatile CRISPR-Cas9-based methods for targeted insertion and replacement of long DNA sequences and genes in the rice genome.
  • To enhance gene editing efficiency for applications in functional genomics and crop trait development.

Main Methods:

  • Utilized chemically modified donor DNA in conjunction with CRISPR-Cas9 for targeted DNA insertion in rice.
  • Employed homology-directed repair, chemically modified donor DNA, and tandem repeats at target sites for gene replacement.
  • Adapted methods previously developed in mammalian cells for plant genome engineering.

Main Results:

  • Successfully inserted sequences up to 2,049 base pairs (bp), including regulatory elements like enhancers and promoters, into the rice genome with 25% efficiency.
  • Achieved gene replacement with up to 130-bp sequences using a homology-directed repair strategy with 6.1% efficiency.

Conclusions:

  • The developed chemically modified donor DNA and CRISPR-Cas9 methods provide versatile tools for inserting long DNA sequences into the rice genome.
  • The gene replacement method offers a new approach for precise editing of target sites in plants.
  • These advancements facilitate functional genomics studies and accelerate trait improvement in crops.