PrimeCap T7 RNA Polymerase (low dsRNA)
Trace amounts of double-stranded RNA (dsRNA) byproduct produced during in vitro transcription can interfere with downstream in vivo administration by triggering unwanted immune responses. PrimeCap T7 RNA Polymerase (low dsRNA) is a modified form of T7 RNA polymerase that combines highly efficient cap-analog-dependent RNA synthesis with extremely low dsRNA generation, making it the ideal choice for researchers engaged in mRNA therapeutic research and development.
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Overview
- Produce in-vitro transcribed RNA with very low dsRNA byproduct that is notably safer for in vivo administration
- Retain the same high yields as T7 RNA polymerase ver.2 (Cat. # 2541A), especially when used with cap analogs such as CleanCap Reagent AG (TriLink Bio Technologies)
- Get improved heat resistance (up to 52°C) compared to T7 RNA polymerase ver.2.0
Produce mRNA with very low dsRNA byproduct—without sacrificing yield
T7 RNA Polymerase ver.2.0 (Cat. #2541A) and PrimeCap T7 RNA Polymerase were used to synthesize mRNA from the Positive Control Template (FLuc) plasmid (sold as part of Cat. #6144). CleanCap Reagent AG (4 mM; TriLink BioTechnologies) and Pyrophosphatase (Cat. #2450A/B) were added to the in vitro transcription reaction. Panel A. mRNA yields of using T7 RNA Polymerase ver.2.0 or PrimeCap T7 RNA Polymerase in a 20 µl reaction volume. Panel B. Percentage of dsRNA per reaction as compared to T7 RNA Polymerase ver.2.0 (WT on y-axis), as determined by a dsRNA ELISA kit.
More Information
Protocol
| RNase-free Water | up to 20 μl |
| 10X T7 RNA Polymerase Buffer | 2 μl |
| ATP, CTP, GTP, UTP | each 10 mM |
| Template DNA | 0.5–2 μg |
| Recombinant RNase Inhibitor ver.2.0 | 20 U |
| Pyrophosphatase (inorganic) | 0.1 U |
| PrimeCap T7 RNA Polymerase | 200 U |
Incubate at 37°C for 1–2 hr.
Applications
- Synthesis of capped RNA using a cap analog
- Synthesis of uncapped RNA for enzymatic capping (note: RNA yield will be 10–40% lower than when using cap analogs)
Source
Escherichia coli carrying a plasmid containing the gene for the modified phage T7 RNA polymerase variant.
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Takara IVTpro mRNA Synthesis System
A complete solution for synthesizing high-quality single-stranded mRNA containing your gene of interest with a cap structure and poly(A) sequence.
Oxidation-resistant Recombinant RNase Inhibitor
The oxidation-resistant Recombinant RNase Inhibitor ver.2.0 inhibits RNAse A and is suitable for use in enzymatic reactions requiring DTT concentrations, such as in vitro transcription, in vitro translation, and RT-PCR.
Inorganic Pyrophosphate
Use our inorganic pyrophosphatase to improve the RNA yield in in vitro transcription reactions.
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