Fig. 1.

Specificity of CsCRUC gRNA spacer design and schematic of CRISPR/Cas9 construct. a Partial sequence alignment of the first exon from CsCRUA, CsCRUB, CsCRUC, and CsCRUD gene families. The CsCRUC gRNA spacer sequences (gRNA512 and gRNA510) used in this study are boxed with the PAM sequence underlined. Numbers indicate nucleotide position from the start codon and shading indicates differences in nucleotide sequence. b Schematic of the CRIPSR/Cas9 construct. Expression of Cas9 is under control of the AtEF1α promoter and encodes a nuclear localization signal (NLS) at the N- and C-termini, plus a 3xFLAG epitope tag. The AtU6–26 promoter drives expression of the CsCRUC sgRNA cassette. Construct is not represented to scale