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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Related Experiment Video

Updated: Jun 19, 2026

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
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Heritable targeted mutagenesis in maize using a designed endonuclease.

Huirong Gao1, Jeff Smith, Meizhu Yang

  • 1Pioneer Hi-Bred International, Research Center, 7300 NW 62nd Avenue, Johnston, IA 50131-1004, USA.

The Plant Journal : for Cell and Molecular Biology
|October 9, 2009
PubMed
Summary
This summary is machine-generated.

Researchers engineered a homing endonuclease to create targeted mutations in maize plants. This molecular tool successfully introduced mutations at the liguleless1 locus in 3% of regenerated plants, demonstrating its utility for genetic modification.

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Area of Science:

  • Plant Molecular Biology
  • Genetic Engineering
  • Biotechnology

Background:

  • Targeted mutagenesis is crucial for understanding gene function and crop improvement.
  • Engineered nucleases offer precise genome editing capabilities.
  • Homing endonucleases, like I-CreI, can be adapted for specific DNA targeting.

Purpose of the Study:

  • To demonstrate targeted mutagenesis in maize using a re-designed single-chain I-CreI endonuclease.
  • To validate the efficacy of this engineered endonuclease for modifying the liguleless1 locus.
  • To assess the frequency and types of mutations induced at the target site.

Main Methods:

  • Design and construction of a single-chain I-CreI endonuclease targeting the LIGULELESS1 (LG1) promoter region.
  • Agrobacterium-mediated transformation of maize immature embryos with the endonuclease gene.
  • Screening of T(0) transgenic plants for mutations at the liguleless1 locus.

Main Results:

  • Mutations were detected at the liguleless1 locus in 3% of T(0) plants.
  • Mutations included monoallelic, biallelic, and chimeric alterations.
  • Short deletions (2-220 bp) were the most common mutations, with some insertions also observed.
  • The engineered endonuclease successfully induced double-strand breaks at the targeted chromosomal locus.

Conclusions:

  • Rational re-design of homing endonucleases yields functional enzymes for targeted mutagenesis.
  • The developed system efficiently introduces targeted mutations in maize without requiring specific selection.
  • Re-designed homing endonucleases are valuable tools for targeted genetic modification in plants.