ArticleProceedings of the National Academy of Sciences of the United States of America2002
Efficient bacterial transcription of DNA nanocircle vectors with optimized single-stranded promoters.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 26 citations in OpenAlex.
- Circular single-stranded DNA as a programmable vector for gene regulation in cell-free protein expression systems.Nature communications · 2024Article
- Direct dsRNA preparation by promoter-free RCT and RNase H cleavage using one circular dsDNA template with a mismatched bubble.RNA (New York, N.Y.) · 2023Article
- Circular single-stranded DNA as switchable vector for gene expression in mammalian cells.Nature communications · 2023Article
- Starting and stopping RNA polymerase III transcription on single-stranded DNA oligonucleotides.RNA (New York, N.Y.) · 2022Article
- The Discovery of Rolling Circle Amplification and Rolling Circle Transcription.Accounts of chemical research · 2016Article
- New insights into the promoterless transcription of DNA coligo templates by RNA polymerase III.Transcription · 2014Article
- Article
- In vivo cloning of artificial DNA nanostructures.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
- Generation of Ribozymes by Rolling Circle Transcription of Promoterless Single-Stranded DNA Circles in Mammalian Cells.Turk biyokimya dergisi = Turkish journal of biochemistry · 2006Article
- A single-stranded promoter for RNA polymerase III.Proceedings of the National Academy of Sciences of the United States of America · 2003Article
- Artificial human telomeres from DNA nanocircle templates.Proceedings of the National Academy of Sciences of the United States of America · 2002Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe experiments aimed at establishing whether circular single-stranded DNAs can form promoters for bacterial transcription from small folded motifs. In vitro selection experiments were carried out on circular 103-nt DNA libraries encoding 40-nt randomized sequences as well as self-processing hammerhead ribozymes. Rounds of rolling circle transcription, reverse transcription-PCR, and recyclization were carried out to optimize transcription efficiency. Sequences were identified that are 80-fold more actively transcribed than the initial library by E. coli RNA polymerase (RNAP). The selected motifs were found to be more active than canonical E. coli promoters in the same context. Experiments also demonstrated that a single-stranded pseudopromoter identified by this selection can be transplanted to other circular DNA contexts and retain transcriptional activity. Results suggest that the promoter is localized in a short ( approximately 40 nt) hairpin, which is smaller than canonical E. coli promoters. To test whether this pseudopromoter was active in bacterial cells, a synthetic DNA nanocircle vector encoding a ribozyme targeted to a site in the marA drug resistance gene was constructed to contain an optimized single-stranded promoter. It is shown that this DNA circle can act as a "Trojan horse" in E. coli, being actively transcribed by the cellular RNAP and producing ribozymes that cleave a sequence in the marA drug resistance gene. The use of optimized single-stranded promoters in combination with synthetic nanocircle DNA vectors represents a potentially useful way to engender the synthesis of biologically active RNAs in living cells.
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