Fig. 1: Preparation of IVT tRNAs, and consequent protein synthesis using IVT recombinant tRNAs or in situ synthesized tRNAs. | Nature Communications

Fig. 1: Preparation of IVT tRNAs, and consequent protein synthesis using IVT recombinant tRNAs or in situ synthesized tRNAs.

From: Continuous in situ synthesis of a complete set of tRNAs sustains steady-state translation in a recombinant cell-free system

Fig. 1

a A schematic of in vitro transcription and a table summarizing the tRNA DNA templates used in this study. Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. b Transcription yield of tRNAs using a linear template containing a T7 class III promoter ϕ6.5 upstream of the wild type tRNA gene (n = 3 replicates). c Comparison of tRNA yield before and after template optimization (n = 3 replicates). d A schematic describing the approach to test the activity of 21 IVT tRNAs in a ΔtRNA PURE system using sfGFP as a reporter. Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. e The abundance of E. coli tRNAs (yellow bars) and the uniform tRNA concentration used (gray dotted line). f Expression of sfGFP using the indicated tRNA source in ΔtRNA PURE system (n = 3 replicates). g Schematic of testing the activity of in situ transcribed tRNAs from linear tRNA templates in ΔtRNA PURE system using sfGFP as a reporter (n = 3 replicates). Created in BioRender. Maerkl, S. (2025) https://BioRender.com/eh7ck4h. h The expression of sfGFP with various input concentrations of linear DNA templates for in situ transcription of tRNAs in the ΔtRNA PURE system (n = 6 for 0 and 42 nM ltDNA, n = 4 for 84 and 252 nM ltDNA, n = 5 for 168 nM ltDNA). Each dot represents a data point from an independent replicate. Bars and error bars represent the mean and standard deviation. Source data are provided as a Source Data file.

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