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Genome Editing in Soybean with CRISPR/Cas9.

Junqi Liu1, Samatha Gunapati1, Nicole T Mihelich1

  • 1Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
PubMed
Summary

This study presents a CRISPR/Cas9 genome editing protocol for soybean, enabling simultaneous targeting of multiple genes. Researchers can now efficiently generate genetically modified soybean plants with mutations in up to three genes.

Keywords:
Agrobacterium-mediated whole plant transformationCRISPR/Cas9Composite plantGradient polyacrylamide gel electrophoresis (PAGE)Hairy root transformation

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Area of Science:

  • Agricultural biotechnology
  • Molecular biology
  • Plant science

Background:

  • CRISPR/Cas9 genome editing technology has advanced rapidly.
  • Applications in diverse plant species, including soybean, are expanding.
  • Platforms for designing and cloning CRISPR targets are available.

Purpose of the Study:

  • To provide an updated protocol for CRISPR/Cas9 gene editing in soybean.
  • To describe methods for targeting single or multiple genes simultaneously.
  • To address limitations in soybean gene editing efficiency.

Main Methods:

  • Utilized two platforms for cloning single and multiplex CRISPR targets.
  • Described reagent delivery methodologies for soybean.
  • Applied protocols to soybean hairy roots, composite plants, and regenerated whole plants.

Main Results:

  • Successfully applied CRISPR/Cas9 technology to soybean.
  • Achieved mutagenesis in up to three target genes simultaneously.
  • Developed protocols addressing critical limiting steps in soybean editing.

Conclusions:

  • The presented protocol enables efficient CRISPR/Cas9 mediated genome editing in soybean.
  • The methods facilitate simultaneous targeting of multiple genes.
  • This work advances the application of genome editing in soybean improvement.