Fig. 4
From: Anti-CRISPR-based biosensors in the yeast S. cerevisiae
β-estradiol sensing circuits. a Upon induction with β-estradiol the chimeric protein LmAcrIIA2-HDB(ER) enters the nucleus and prevents dSpCas9-Mxi1:gRNA from binding Tsynth8_pCYC1noTATA promoter. Hence, the overall effect of β-estradiol is to activate mRNA transcription from Tsynth8_pCYC1noTATA, as symbolized by the green arrow. The dashed red line indicates that, in its ground-state configuration, LmAcrIIA2-HDB(ER) is unable to interact with dSpCas9-Mxi1:gRNA due to the binding of Hsp90 to HBD(ER). b-c Titration curves. The fluorescence gain of the A2_HBD-RGR (b) and A2_HBD-pSNR52i (c) upon induction with β-estradiol is estimated for 11 different hormone concentrations. Each fluorescence value was obtained as the mean of four independent measurements (3 replicates only were available for A2_HBD-RGR at 31.25 nM and A2_HBD-pSNR52i at 2 μM). The standard deviations of the mean values of relative fluorescence were calculated via the error propagation formula