Figure 2 | Scientific Reports

Figure 2

From: Highly efficient heritable targeted deletions of gene clusters and non-coding regulatory regions in Arabidopsis using CRISPR/Cas9

Figure 2

CRISPR/Cas9 targeted deletions of the ISU1 locus and genetic complementation with an ISU1-YFP reporter. (A) Schematic representation of the ISU1 locus and the Cas9 binding sites. (B) PCR analysis of the ISU1 locus with oligonucleotides flanking the deletion site. The expected PCR size containing a deletion is 500 bp. Primers JD157 with JD158 and JD125 with JD126 were used for the ISU1 deletion and At5g60390 control, respectively. (C) Sanger sequencing results of four independent deletions (T1) (sgRNA sequence in red and PAM sequence in blue). (D) Defects in early embryo development associated with isu1-d1 and isu1–1 (GK424D02) mutations. Scale bar: 50 μm. in, integuments, en, endosperm, em, embryo, su, suspensor (E) Confocal microscopy images of roots from two independent ISU1-YFP transgenic lines that complement the isu1-d1 mutant. Scale bar: 50 μm. (F) Genotype analysis of plants from two independent isu1-d1/ISU1-YFP complemented lines. Deletions of the ISU1 locus was determined by PCR using oligonucleotides flanking the deletion site. The expected PCR product size for the deletion is 500 bp and the expected PCR size for the wild-type ISU1 allele is 600 bp. Primers JD157 with JD159 and JD157 with JD158 were used for the ISU1 wild-type allele and ISU1 deletion allele, respectively. W, wild-type; H, homozygous; h, heterozygous.

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