Skip to main content
Fig. 1 | Plant Methods

Fig. 1

From: Targeted gene deletion with SpCas9 and multiple guide RNAs in Arabidopsis thaliana: four are better than two

Fig. 1

Constructs and target selection for deletion of the AtWRKY30 locus. a Recipient vectors pDGE1108 and pDGE1109. The FAST (Fluorescence Accumulating Seed Technology) marker allows positive and negative selection of transgenics. The intron-optimized zCas9i gene is under control of the Arabidopsis RPS5a (Ribosomal Particle S5a) promoter and a chimeric terminator (“t-triple “: t35S-tNbACT-Rb7MAR; [12]). b Scheme of the WRKY30 locus and selected target sites (guide RNAs). Three pairs of target sites (triangles; inner—pink/light pink, middle—blue/light blue, outer—green/light green) were selected and corresponding guide RNAs designed. Arrowheads represent binding sites of oligonucleotides used for PCR genotyping. c Scheme of multiplex editing vectors assembled for WRKY30 editing and containing or not TREX2. Constructs contain cassettes for expression of pairs of guide RNAs, pairs of pairs, or all six guide RNAs

Back to article page

Plant Methods

ISSN: 1746-4811