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Fig. 4 | Plant Methods

Fig. 4

From: Combined fluorescent seed selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants

Fig. 4

CRISPR/Cas T-DNA vectors for further modification and improvement. a Schematic representation of T-DNA vector (pRU319) containing KpnI, SgsI and HindIII restrictions sites flanking the promoter, Cas gene and selection cassette, accordingly. b FastCyan seed selection based on seed-specific pOLE::OLE1-mTurquoise2 expression. Yellow arrows indicate the positive fluorescent seeds. c Schematics of SGN3 gene showing the positions of the chosen sgRNAs. d Basic Fuchsin (gray) staining of lignin-based Casparian strips in the isolated T1 generation biallelic sgn3 mutants generated using pEC1.2::Cas9 and pEC1.2::zCas9i constructs. e Segregation analysis in T1 generation in pEC1.2::Cas9, pEC1.2::zCas9i, PcUBi4-2::Cas9 and PcUBi4-2::zCas9i lines. f Segregation analysis of T2 generation in pEC1.2::Cas9 and pEC1.2::zCas9i lines. Scale bars  = 100 μm (B) and in 25 μm (D)

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