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. 2018 Sep 5;4(9):eaat6086. doi: 10.1126/sciadv.aat6086

Fig. 1. Genome editing of MeGBSS and MePTST1.

Fig. 1

(A) Model for association between GBSS, PTST1, and starch granules as proposed by Seung et al. (35). Image recreated with permission. (B) Binary expression construct containing hptII (hygromycin B resistance) for plant selection; Cas9 codon optimized for cassava usage (pcoCas9) and fused to the eGFP reporter gene and directed to the nucleus via a nucleoplasmin nuclear localization signal (NLS); transcription terminated by sequence from a heat shock protein (tHSP) (56). A synthetic pU6 promoter [(psynU6; 10)] used to drive expression of the protospacer sequence [single guide RNA (sgRNA)] and synthetic scaffold (9) to generate the desired sgRNA. AtFT for early flowering is constitutively expressed by the CaMV35S promoter and terminated by nopaline synthase sequence (tNOS). Binary expression constructs named pCas9-sgGBSS-FT and pCas9-sgPTST-FT. Left and right borders (LB and RB) are shown. Diagram not to scale. (C) Gene maps of MeGBSS and MePTST1. Exons are depicted as blocks, and regions encoding functional domains are shaded. The target sites for the sgRNAs are indicated. Scale bars are shown.