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Fig. 1 | Plant Methods

Fig. 1

From: New improvements in grapevine genome editing: high efficiency biallelic homozygous knock-out from regenerated plantlets by using an optimized zCas9i

Fig. 1

Schematic illustration of target gene and different plasmid constructions. (a) Illustration of the target gene belonging to LysM receptor-like kinase family (Vitvi05g00623) with the location in red of the sgRNA1, 2 and 3 in exon1. (b) Detailed sgRNA sequences with the Protospacer Adjacent Motif (PAM) underlined and the GC content. (c) Plasmid construction for grapevine embryogenic cells transformation. Both constructions are constituted of one cassette with NPTII gene under the control of NOS promoter for antibiotic selection, one cassette expressing the DsRed2 gene under the control of 35 S promoter for fluorescent selection and a multiplex of 3 sgRNAs under the control of U6-26 promoter from A. thaliana. Constructions differ by the nucleotide sequence encoding Cas9 gene with integration of 13 introns for zCas9i and by the use of two promoters controlling Cas9 gene expression

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Plant Methods

ISSN: 1746-4811