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Fig. 1 | Plant Methods

Fig. 1

From: A simple and efficient CRISPR/Cas9 platform for induction of single and multiple, heritable mutations in barley (Hordeum vulgare L.)

Fig. 1

Schematic description of RNA-guided Cas9 constructs designed for genome editing. a Structure of the binary vector plasmid based on pBract211 used to deliver Cas9:sgRNA components into barley plants. The Gateway cloning site is replaced by the sgRNA cassette; P-ZmUbi, maize ubiquitin promoter; Cas9-int, synthetic gene of nuclease Cas9 with an intron and nuclear localization signal; nos, nopaline synthase terminator; P-35S, CaMV35S promoter; Hyg-int, hygromycin resistance gene; kmR, kanamycin resistance gene. b Structure of the pCR8/GW/TOPO-sgRNA vector used for assembling the sgRNA or PTG constructs. Either oligo duplex for simplex editing or polycistronic tRNA-gRNA (PTG) for multiplex editing can be cloned between the U6 promoter and gRNA scaffold using BsaI-generated overhangs. c Structure of the polycistronic tRNA-gRNA unit with two spacers used for multiplex editing

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Plant Methods

ISSN: 1746-4811