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Genome Editing in Soybean with CRISPR/Cas9

  • Junqi Liu
  • Samatha Gunapati
  • Nicole T. Mihelich
  • Adrian O. Stec
  • Jean-Michel Michno
  • Robert M. StuparEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)

Abstract

CRISPR/Cas9 mediated genome editing technology has experienced rapid advances in recent years and has been applied to a wide variety of plant species, including soybean. Several platforms have been developed for designing and cloning of single CRISPR targets or multiple targets in a single destination vector. This chapter provides an updated working protocol for applying CRISPR/Cas9 technology to target a single gene or multiple genes simultaneously in soybean. We describe two platforms for cloning single CRISPR targets and multiplexing targets, respectively, and reagent delivery methodologies. The protocols address crucial limiting steps that can limit CRISPR editing in soybean hairy roots, composite plants, and tissue culture-based regenerated whole plants. To date, transgenic soybean plants with mutagenesis in up to three target genes have been obtained with this procedure.

Key words

CRISPR/Cas9 Hairy root transformation Composite plant Agrobacterium-mediated whole plant transformation Gradient polyacrylamide gel electrophoresis (PAGE) 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Junqi Liu
    • 1
  • Samatha Gunapati
    • 1
  • Nicole T. Mihelich
    • 1
  • Adrian O. Stec
    • 1
  • Jean-Michel Michno
    • 1
  • Robert M. Stupar
    • 1
    Email author
  1. 1.Department of Agronomy and Plant GeneticsUniversity of MinnesotaSt. PaulUSA

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