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

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

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Soybeans
Organism Cloning
Genes
Technology
Plant Roots
Genetically Modified Plants
Mutagenesis
Gene Editing

Keywords

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

PubMed: MeSH publication types

  • Journal Article

Cite this

Liu, J., Gunapati, S., Mihelich, N. T., Stec, A. O., Michno, J. M., & Stupar, R. M. (2019). Genome editing in soybean with CRISPR/Cas9. In Methods in Molecular Biology (pp. 217-234). (Methods in Molecular Biology; Vol. 1917). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8991-1_16

Genome editing in soybean with CRISPR/Cas9. / Liu, Junqi; Gunapati, Samatha; Mihelich, Nicole T; Stec, Adrian O.; Michno, Jean Michel; Stupar, Robert M.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 217-234 (Methods in Molecular Biology; Vol. 1917).

Research output: Chapter in Book/Report/Conference proceedingChapter

Liu, J, Gunapati, S, Mihelich, NT, Stec, AO, Michno, JM & Stupar, RM 2019, Genome editing in soybean with CRISPR/Cas9. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1917, Humana Press Inc., pp. 217-234. https://doi.org/10.1007/978-1-4939-8991-1_16
Liu J, Gunapati S, Mihelich NT, Stec AO, Michno JM, Stupar RM. Genome editing in soybean with CRISPR/Cas9. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 217-234. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8991-1_16
Liu, Junqi ; Gunapati, Samatha ; Mihelich, Nicole T ; Stec, Adrian O. ; Michno, Jean Michel ; Stupar, Robert M. / Genome editing in soybean with CRISPR/Cas9. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 217-234 (Methods in Molecular Biology).
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