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Fig. 2 | Microbial Cell Factories

Fig. 2

From: CRISPR/dCas9-mediated transcriptional improvement of the biosynthetic gene cluster for the epothilone production in Myxococcus xanthus

Fig. 2

Design of the sgRNA and the mxdCas9-activator fusion for RNA-guided transcriptional activation. a A physical map of the promoter region for the epothilone biosynthetic gene cluster, showing the places of five designed spacers. The epothilone genes are shown in green arrow, and the two TSSs are shown with red lines. The different spacer sequences are shown in bars with different colors. b The molecular structures of epothilones A and B. c Design of sgRNA scalfold, containing a 20-nt specific targeting sequence, the tracrRNA scaffold and a U6 terminator. d The ω subunit protein was designed to fuse to the C-terminal of the mxdCas9 with a linker. This fusion gene is under the control of T7A1 promoter. The two mutation sites (D10A and H840A) are shown

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