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Related Experiment Video

Updated: Dec 21, 2025

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
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Complex Trait Loci in Maize Enabled by CRISPR-Cas9 Mediated Gene Insertion.

Huirong Gao1, Jasdeep Mutti1, Joshua K Young1

  • 1Research and Development, Corteva Agriscience, Johnston, IA, United States.

Frontiers in Plant Science
|May 21, 2020
PubMed
Summary

Researchers developed a complex trait locus (CTL) using CRISPR-Cas9 to efficiently stack multiple biotech traits in maize. This innovation simplifies the development of multi-trait maize hybrids, reducing costs and complexity.

Keywords:
CRISPR-Cas9complex trait locigene expressiongene targetgenetic crossingmaizerecombinase-mediated cassette exchangetrait stack

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Area of Science:

  • Agricultural Science
  • Genetics and Genomics
  • Biotechnology

Background:

  • Modern maize hybrids incorporate numerous biotech and native traits.
  • Current methods for inserting multiple traits are inefficient, costly, and complex due to random genome insertion.
  • Developing maize with stacked traits is a significant challenge in crop improvement.

Purpose of the Study:

  • To introduce a novel method for efficient trait stacking in maize using CRISPR-Cas9 technology.
  • To develop a complex trait locus (CTL) for targeted insertion of multiple genes.
  • To overcome the limitations of random gene insertion in creating multi-trait maize hybrids.

Main Methods:

  • Utilized CRISPR-Cas9 gene editing to engineer a complex trait locus (CTL) in maize.
  • Generated individual maize lines with site-specific insertion landing pads (SSILPs) at preselected sites within the CTL.
  • Employed recombinase-mediated cassette exchange for efficient transgene insertion into SSILPs.

Main Results:

  • Successfully generated maize lines with SSILPs at targeted sites within the CTL.
  • Demonstrated consistent transgene expression from selected insertion sites.
  • Confirmed that SSILP insertion did not negatively impact grain yield.
  • Showcased the ability to combine two traits at different CTL sites via genetic recombination.

Conclusions:

  • CTL technology offers a streamlined and efficient approach for developing multi-trait maize hybrids.
  • This method significantly reduces the complexity and cost associated with trait stacking.
  • CTL represents a major advancement in the genetic engineering of maize for improved crop performance.