JEDLIČKOVÁ, Veronika, Kateřina MÁCOVÁ, Marie ŠTEFKOVÁ, Jan BUTULA, Jana ZEMÁNKOVÁ, Marek SEDLÁČEK and Helene ROBERT BOISIVON. Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus). Frontiers in Plant Science. SWITZERLAND: Frontiers Research Foundation, 2022, vol. 13, August, p. 1-15. ISSN 1664-462X. Available from: https://dx.doi.org/10.3389/fpls.2022.919290.
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Basic information
Original name Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus)
Authors JEDLIČKOVÁ, Veronika (203 Czech Republic, belonging to the institution), Kateřina MÁCOVÁ (203 Czech Republic, belonging to the institution), Marie ŠTEFKOVÁ (203 Czech Republic, belonging to the institution), Jan BUTULA (203 Czech Republic, belonging to the institution), Jana ZEMÁNKOVÁ (203 Czech Republic, belonging to the institution), Marek SEDLÁČEK (203 Czech Republic, belonging to the institution) and Helene ROBERT BOISIVON (250 France, guarantor, belonging to the institution).
Edition Frontiers in Plant Science, SWITZERLAND, Frontiers Research Foundation, 2022, 1664-462X.
Other information
Original language English
Type of outcome Article in a journal
Country of publisher Switzerland
Confidentiality degree is not subject to a state or trade secret
WWW URL
RIV identification code RIV/00216224:14740/22:00126354
Organization Středoevropský technologický institut – Repository – Repository
Doi http://dx.doi.org/10.3389/fpls.2022.919290
UT WoS 000844031100001
Keywords in English hairy root; CRISPR/Cas9; genome-editing; Brassica napus; TAA1
Links EF16_026/0008446, research and development project. GA19-05200S, research and development project. LM2018129, research and development project.
Changed by Changed by: RNDr. Daniel Jakubík, učo 139797. Changed: 26/8/2023 04:03.
Abstract
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.
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