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Fig. 3 | Plant Methods

Fig. 3

From: Ribozyme-mediated CRISPR/Cas9 gene editing in pyrethrum (Tanacetum cinerariifolium) hairy roots using a RNA polymerase II-dependent promoter

Fig. 3

Analysis of gene editing of transgenic hairy roots 1 month after subculture. a Control group (CK). b Line A. c Line B. d Detection of the RGR sequence by PCR. M, marker; +, ribozyme-based CRISPR/Cas9 vector; −, CK; 1, line A; 2, line B. e PCR amplification followed by restriction enzyme digestion detection of target sites: M, marker; 1−, CK TcEbFS amplicon; 1+, CK TcEbFS fragments following digestion with by BstbI; 2−, Line A TcEbFS amplicon; 2+, Line A TcEbFS fragments following digestion with BstbI; 3−, Line B TcEbFS amplicon; 3+, Line B TcEbFS fragments following digestion with BstbI. f DNA sequencing confirming TcEbFS gene mutations in pyrethrum hairy roots. The net length of insertions and/or deletions are presented in the column to the left. Numbers in parentheses indicate the number of clones. g Diagram showing the TcEbFS genomic locus with the sgRNA targeting site and primer locations (arrows). Scale bars in ac, 1.0 cm

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