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| Item Type: | Article |
|---|---|
| Title: | Maximizing transcription of nucleic acids with efficient T7 promoters |
| Creators Name: | Conrad, T., Plumbom, I., Alcobendas, M., Vidal, R. and Sauer, S. |
| Abstract: | In vitro transcription using T7 bacteriophage polymerase is widely used in molecular biology. Here, we use 5'RACE-Seq to screen a randomized initially transcribed region of the T7 promoter for cross-talk with transcriptional activity. We reveal that sequences from position +4 to +8 downstream of the transcription start site affect T7 promoter activity over a 5-fold range, and identify promoter variants with significantly enhanced transcriptional output that increase the yield of in vitro transcription reactions across a wide range of template concentrations. We furthermore introduce CEL-Seq(+) , which uses an optimized T7 promoter to amplify cDNA for single-cell RNA-Sequencing. CEL-Seq(+) facilitates scRNA-Seq library preparation, and substantially increases library complexity and the number of expressed genes detected per cell, highlighting a particular value of optimized T7 promoters in bioanalytical applications. |
| Keywords: | AT Rich Sequence, Bacteriophage T7, Base Sequence, Genetic Promoter Regions, Genetic Templates, Genetic Transcription, Nucleic Acids, RNA-Seq, Single-Cell Analysis |
| Source: | Communications Biology |
| ISSN: | 2399-3642 |
| Publisher: | Springer Nature |
| Volume: | 3 |
| Number: | 1 |
| Page Range: | 439 |
| Date: | 14 August 2020 |
| Official Publication: | https://doi.org/10.1038/s42003-020-01167-x |
| PubMed: | View item in PubMed |
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