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. 2023 Aug 10;13:12961. doi: 10.1038/s41598-023-39777-0

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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In vitro transcription of DNA nanoparticles. Representative non-denaturing agarose gel electrophoresis results of in vitro transcription reactions. M: molecular marker; ss: ssT7EGFP; ds: dsT7GFP; Lp: LpCMV-T7-EGFP (linearized GFP plasmid); PO: T7GHL PO; HS: T7GHL HS; FS: T7GHL FS; BP: T7GHL BP; -T7: T7GHL-T7. LpCMV-T7-EGFP and dsT7EGFP were positive controls. ssT7EGFP, T7GHL BP, and T7GHL -T7 were putative negative controls. The bands at approximately 0.9 kb, 1.1 kb, and 1.3 kb represent GFP RNA. The 0.9 kb band represents transcripts that terminate at a T7 RNA polymerase termination/stall sequence, known to allow a significant amount of read-through^32,33. The 1.1 kb and 1.3 kb RNA products represent read-through transcripts, the latter’s greater length resulting from the linearized plasmid being 200 bp longer than the PCR-generated linear duplex. The bands of approximately 2.5 kb for most of the samples and 1.5 kb in the T7GHL -T7 lane result from the presence of residual DNA templates.