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Fig. 1 | Rice

Fig. 1

From: Targeted Deletion of the First Intron of the Wxb Allele via CRISPR/Cas9 Significantly Increases Grain Amylose Content in Rice

Fig. 1

CRISPR/Ca9-mediated deletion of the Wx 1st intron. A Position and sequence of the two gRNA target sites on the genomic regions of the Wx gene. Introns are shown as lines. Exons are shown as boxes. PAM motifs (CCN) of two targeted sequences are underlined. Two primer pairs, JC-F/R used for the detection of large fragment deletions and RT-F/R used for Wx gene expression analysis via RT–qPCR, were marked at their appropriate locations. B Schematic diagram of the editing vector pZZT477’s T-DNA structure. LB, T-DNA left border; RB, T-DNA right border; 35S, CaMV35s promoter; CP4, Agrobacterium tumefaciens strain CP4 EPSP (5-enolpyruvylshikimate-3-phosphate synthase) gene; Ubi4, sugarcane ubiquitin 4 gene promoter; Cas9, CRISPR/Cas9 gene with codon optimized for expression in rice; U3, U6, rice U3 and U6 snRNA promoter. C Detection of mutations in the first intron of the Wx gene via PCR assay in the T0 generation. D Sequencing results of the first intron-deleted mutant lines. The PAM motifs are underlined, target sequences are highlighted in red, and dotted lines indicate deletions. The WT indicates the wild type, and M1, M2, and M3 are different homozygous lines in the T1 generation

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