Fig. 7: PsiR-mediated gene expression repression and dynamic metabolic regulation. | Nature Communications

Fig. 7: PsiR-mediated gene expression repression and dynamic metabolic regulation.

From: Computational design of allulose-responsive biosensor toolbox for auto-inducible protein expression and CRISPRi mediated dynamic metabolic regulation

Fig. 7: PsiR-mediated gene expression repression and dynamic metabolic regulation.

a The schematic diagram of the designed allulose-triggered CRISPR interference (ATCi) for regulating gene expression. b The verification of our designed ATCi for repressing eGFP and mRFP1 expression in C. glutamicum chromosome. G-rA: CgdC-M2-GR (pXMJ19-crRNA-eGFP218); G-rB: CgdC-M2-GR (pXMJ19-crRNA-eGFP663); R-rA: CgdC-M2-GR (pXMJ19-crRNA-mRFP169); R-rB: CgdC-M2-GR (pXMJ19-crRNA-mRFP359); G-rA+R-rA: CgdC-M2-GR (pXMJ19-crRNA-eGFP218-mRFP169). Also, the confocal images of strain CgdC-M2-GR (pXMJ19-crRNA-eGFP218) from the green channel (515–564 nm) under 0 and 5 mM allulose were shown. c Metabolic engineering strategy for engineering the allulose synthetic cell factory. Glc Glucose, G6P glucose-6-phosphate, F6P fructose-6-phosphate, A6P allulose-6-phosphate, 1,6-FBP d-fructose 1,6-bisphosphate, 6PG glucono-1,5-lactone 6-phosphate, BsglcK glcK from Bacillus subtilis, Bspgi pgi from Bacillus subtilis. d–g The cell growth (OD600), glucose consumption, allulose synthesis, and lactate synthesis for the strains CgAll1, CgAll2, and CgAll3. Data are presented as mean values +/−SD (n = 3 independent experiments). Source data for (b–d–g) are provided as a Source Data file.

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