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. Author manuscript; available in PMC: 2021 Aug 13.
Published in final edited form as: ACS Synth Biol. 2021 Feb 12;10(3):459–465. doi: 10.1021/acssynbio.0c00402

Figure 1. Cell-free gene repression using small antisense DNA.

Figure 1

(A) Schematic overview of the working principle of the cf-sDNA technique. Following in vitro transcription of mRNA, sDNA already present in solution hybridizes with the targeted mRNA at the ribosome binding site and downstream sequence, which inhibits translational initiation and elongation. (B)+(C) Repression of YPet expression using different concentrations of sDNA as determined from YPet fluorescence. In (B) mRNA is transcribed from a T7 promoter while in (C) it is produced from an E.coli promoter (J23106). The ON/OFF YPet fluorescence is the ratio between the fluorescence of a negative control (0 μM sDNA, YPet fluorescence “ON”) and the fluorescence determined in samples with sDNA (YPet fluorescence “OFF”). Fluorescence values used are biological triplicates of fluorescence end levels after 13-14 h of incubation in the cell-free expression system. The given uncertainties are S.E., but they are hardly visible due to their small values.