Publication details

Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus)

Investor logo
Investor logo
Authors

JEDLIČKOVÁ Veronika MÁCOVÁ Kateřina ŠTEFKOVÁ Marie BUTULA Jan ZEMÁNKOVÁ Jana SEDLÁČEK Marek ROBERT BOISIVON Helene

Year of publication 2022
Type Article in Periodical
Magazine / Source FRONTIERS IN PLANT SCIENCE
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://doi.org/10.3389/fpls.2022.919290
Doi http://dx.doi.org/10.3389/fpls.2022.919290
Keywords hairy root, CRISPR/Cas9, genome-editing, Brassica napus, TAA1
Attached files
Description Our study examined the mutation efficiency of the CRISPR/Cas9 method for tryptophan aminotransferase BnaTAA1 genes involved in the auxin biosynthesis pathway. We made nine CRISPR/Cas9 constructs with various promoters driving the expression of a Cas9 from Staphylococcus aureus (SaCas9) or a plant-codon-optimized Streptococcus pyogenes Cas9 (pcoCas9). We developed a fast and efficient system for evaluating the variety and frequency of mutations caused by each construct using Brassica napus hairy roots. We showed that pcoCas9 is more efficient in mutating the targeted loci than SaCas9 and the presence of the NLS signal enhanced the chance of mutagenesis by 25%. The mutations were studied further in regenerated lines, and we determined the BnaTAA1 gene expression and heritability of the gene modifications in transgenic plants. Hairy root transformation combined with CRISPR/Cas9-mediated gene editing represents a fast and straightforward system for studying target gene function in the important oilseed crop B. napus.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.

More info