Extended Data Fig. 3: The transcription–translation combination switch-mediated protein expression.
From: Genetic-code-expanded cell-based therapy for treating diabetes in mice
a, Schematic showing the design of the transcription-translation combination switch. A ncAA could trigger translation of tetracycline-controlled transactivator (tTA) bearing an ectopic amber codon, then Dox could stop SEAP transcription by regulating the activity of the tTA. b, The translation-transcription combination switch consists of triple plasmids, encoding an OmeYRS/tRNA pair, tTA with an amber codon, and wild-type SEAP. c, Exploration of the impact on amber codon position on the expression of a POI (transactivator tTA) in HEK293T cells. OmeYRS/tRNA pair was used for OmeY incorporation. SEAP levels in culture supernatants were measured 48 h after OmeY or Dox treatment. Data are presented as the mean ± SD; n = 3 independent samples. The signal-to-noise ratio is indicated above the bars. Red rectangle represents the final construct used in subsequent studies. d, Constructs of translation-transcription combination switch using bacterial TyrRS/tRNA, and fluorescence micrographs of designer cells cultivated within or without OmeY. Each experiment was repeated three times independently with similar results. e, Constructs of translation-transcription combination switch using archaea PylRS/tRNA, and fluorescence micrographs of designer cells cultivated within or without BocK. Each experiment was repeated three times independently with similar results.
