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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Transcriptional reprogramming in yeast using dCas9 and combinatorial gRNA strategies

Fig. 1

Comparing two dCas9 systems for transcriptional regulation. a The anhydro-tetracyclin (aTc) inducible system with gRNA expression controlled by TetO reprograms transcriptional expression with dCas9 directly fused to either VPR or Mxi1 anchoring to promoters of genes of interest (GOI). b The constitutive dCas9 and scRNA expression system regulates transcription through orthogonal gRNA scaffold extensions that recruit endogenously transcribed Mxi1 or VPR. Two identical effectors can be recruited per scRNA. Introducing dCas9 and scRNA(s) promote the onset of this system. For both the inducible and the constitutive system Mxi1 (red) is used for repression and VPR (green) for activation. c Benchmarking the inducible and the constitutive system. BioLector data from time-point 24 h are shown for both systems as relative MFI compared to controls. Control levels are shown in black, repression in light grey and activation in dark grey. Results are presented as GFP/OD from targeting the GFP-fused promoters HMG1 at position TSS-128 and OLE1 at position TSS-381 in both systems. MFI values are shown as mean ± S.D. from three (n = 3) biological replicate experiments

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