Přehled o publikaci
2022
Hairy root transformation system as a tool for CRISPR/Cas9-directed genome editing in oilseed rape (Brassica napus)
JEDLIČKOVÁ, Veronika; Kateřina MÁCOVÁ; Marie ŠTEFKOVÁ; Jan BUTULA; Jana ZEMÁNKOVÁ et al.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; Kateřina MÁCOVÁ; Marie ŠTEFKOVÁ; Jan BUTULA; Jana ZEMÁNKOVÁ; Marek SEDLÁČEK and Helene ROBERT BOISIVON
Edition
Frontiers in Plant Science, SWITZERLAND, Frontiers Research Foundation, 2022, 1664-462X
Other information
Language
English
Type of outcome
Article in a journal
Country of publisher
Switzerland
Confidentiality degree
is not subject to a state or trade secret
References:
Marked to be transferred to RIV
Yes
RIV identification code
RIV/00216224:14740/22:00126354
Organization
Středoevropský technologický institut – Repository – Repository
UT WoS
EID Scopus
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. Czech-BioImaging II, large research infrastructures.
Changed: 9/10/2024 00:50, RNDr. Daniel Jakubík
Abstract
In the original language
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.
