Genome editing in soybean with CRISPR/Cas9

Junqi Liu, Samatha Gunapati, Nicole T. Mihelich, Adrian O. Stec, Jean Michel Michno, Robert M. Stupar

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages217-234
Number of pages18
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1917
ISSN (Print)1064-3745

Keywords

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

PubMed: MeSH publication types

  • Journal Article

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