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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@article{51287, author = {Jedličková, Veronika and Mácová, Kateřina and Štefková, Marie and Butula, Jan and Zemánková, Jana and Sedláček, Marek and Robert Boisivon, Helene}, article_location = {SWITZERLAND}, article_number = {August}, doi = {http://dx.doi.org/10.3389/fpls.2022.919290}, keywords = {hairy root; CRISPR/Cas9; genome-editing; Brassica napus; TAA1}, language = {eng}, issn = {1664-462X}, journal = {Frontiers in Plant Science}, title = {Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus)}, url = {https://doi.org/10.3389/fpls.2022.919290}, volume = {13}, year = {2022} }
TY - JOUR ID - 51287 AU - Jedličková, Veronika - Mácová, Kateřina - Štefková, Marie - Butula, Jan - Zemánková, Jana - Sedláček, Marek - Robert Boisivon, Helene PY - 2022 TI - Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus) JF - Frontiers in Plant Science VL - 13 IS - August SP - 1-15 EP - 1-15 PB - Frontiers Research Foundation SN - 1664-462X KW - hairy root KW - CRISPR/Cas9 KW - genome-editing KW - Brassica napus KW - TAA1 UR - https://doi.org/10.3389/fpls.2022.919290 N2 - 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. ER -
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). \textit{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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