Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa, using CRISPR/Cas9 gene editing

Wen Zhi Jiang, Isabelle M. Henry, Peter G. Lynagh, Luca Comai, Edgar B. Cahoon, Donald P. Weeks

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

The CRISPR/Cas9 nuclease system is a powerful and flexible tool for genome editing, and novel applications of this system are being developed rapidly. Here, we used CRISPR/Cas9 to target the FAD2 gene in Arabidopsis thaliana and in the closely related emerging oil seed plant, Camelina sativa, with the goal of improving seed oil composition. We successfully obtained Camelina seeds in which oleic acid content was increased from 16% to over 50% of the fatty acid composition. These increases were associated with significant decreases in the less desirable polyunsaturated fatty acids, linoleic acid (i.e. a decrease from ~16% to <4%) and linolenic acid (a decrease from ~35% to <10%). These changes result in oils that are superior on multiple levels: they are healthier, more oxidatively stable and better suited for production of certain commercial chemicals, including biofuels. As expected, A. thaliana T2 and T3 generation seeds exhibiting these types of altered fatty acid profiles were homozygous for disrupted FAD2 alleles. In the allohexaploid, Camelina, guide RNAs were designed that simultaneously targeted all three homoeologous FAD2 genes. This strategy that significantly enhanced oil composition in T3 and T4 generation Camelina seeds was associated with a combination of germ-line mutations and somatic cell mutations in FAD2 genes in each of the three Camelina subgenomes.

Original languageEnglish (US)
Pages (from-to)648-657
Number of pages10
JournalPlant Biotechnology Journal
Volume15
Issue number5
DOIs
StatePublished - May 2017

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Camelina
Camelina sativa
Seeds
Fatty Acids
fatty acid composition
seeds
Oils
genes
mutation
oils
Guide RNA
oilseed crops
nucleases
Genes
linolenic acid
biofuels
seed oils
somatic cells
alpha-Linolenic Acid

Keywords

  • CRISPR/Cas9
  • Camelina sativa
  • allohexaploid
  • fatty acid composition
  • gene editing
  • oleic acid

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Plant Science

Cite this

Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa, using CRISPR/Cas9 gene editing. / Jiang, Wen Zhi; Henry, Isabelle M.; Lynagh, Peter G.; Comai, Luca; Cahoon, Edgar B.; Weeks, Donald P.

In: Plant Biotechnology Journal, Vol. 15, No. 5, 05.2017, p. 648-657.

Research output: Contribution to journalArticle

Jiang, Wen Zhi ; Henry, Isabelle M. ; Lynagh, Peter G. ; Comai, Luca ; Cahoon, Edgar B. ; Weeks, Donald P. / Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa, using CRISPR/Cas9 gene editing. In: Plant Biotechnology Journal. 2017 ; Vol. 15, No. 5. pp. 648-657.
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