The T7 mScript™ Complete Standard mRNA Production System provides all enzymes and enzyme-related reagents for making canonical uridine-containing, 5'-capped, 3'-polyadenylated mRNA with greatly reduced double-stranded RNA (dsRNA) content, lower than what can be achieved with special T7 RNA polymerase mutants. The kit includes modules for (i) in vitro transcription of linear double-stranded DNA templates using the T7 mScript™ Enzyme Solution and the canonical nucleotides ATP, CTP, GTP and UTP, (ii) enzymatic capping of the RNA using the ScriptCap™ Cap 1 Capping System (contains both ScriptCap™ Capping Enzyme and 2'-O-Methyltransferase) for making mRNA with a Cap 1 cap structure, (iii) A-Plus™ Poly(A) Polymerase for adding a 3'-poly(A) tail, (iv) enzymatic removal of dsRNA content using the Min-Immune™ Gold dsRNA Removal system and (v) 5 M NH₄OAc as a convenient RNA/mRNA purification method.

Post-transfection, capped and tailed mRNA has increased stability and translation efficiency in most eukaryotic cell lines. The mScript™ System improves upon existing capping methods by ensuring virtually 100% transcript capping, all caps in the proper orientation and the ability to produce large amounts of capped RNA at a reasonable cost. The mScript™ System uses post-transcriptional poly(A) tailing allowing the user to define their desired tail length. In this way, poly(A) tails can be generated much longer than is possible co-transcriptionally using a template-encoded tail. mRNAs with longer poly(A) tails are expressed for longer periods of time in cells. Additionally, the removal of dsRNA from mRNA preparations has been shown to be essential for reducing the innate immunogenic response to the mRNA in cells.^1-4 Alternative dsRNA removal methods, such as reverse-phase HPLC⁵, hydroxy­apatite chromatography⁶ and cellulose chromatography⁷, are associated with high capital costs as well as reduced final product yields. The Min-Immune™ Gold dsRNA Removal system provides an easy-to-use method for removing the dsRNA content of mRNA preps without a reduction of the single-stranded mRNA yield. mScript™ Complete U-mRNA can be used in many transfection and microinjection experiments as well as in vitro translation systems. However, it has been shown that N1meΨ-mRNAs and Ψ-mRNAs are translated into protein at higher levels and induce lower innate immune responses in human and other mammalian cells that express various RNA sensors compared to corresponding unmodified mRNAs.^8-13 CELLSCRIPT™ also offers the INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System and INCOGNITO™ T7 mScript™ Complete Ψ-mRNA Production System for making these types of mRNAs, respectively.

Materials Supplied

Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.

T7 Control Template DNA: Is a linearized 4.1 kb plasmid that contains a T7 promoter followed by a phage lambda dsDNA insert that encodes a 1,375 base runoff transcript. The Control Template DNA is provided at a concentration of 0.5 µg/µl in T₁₀E₁ Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA).

Materials Required, but not Supplied

Optional Materials

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Manual

SDS

These are both links to the manufacturer's page

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1. Karikó, K. et al., (2011) Nucleic Acids Res. 39, e142.
2. Schlee, M., Hartmann, G., (2016) Nat. Rev. Immunol. 16, 566.
3. Pardi, N. et al., (2018) Nat. Rev. Drug Discov. 17, 261.
4. Mu, X., Hur, S., (2021) Acc. Chem. Res. 54, 4012.
5. Weissman, D. et al., (2013) Methods Mol. Biol. 969, 43.
6. Kalmakoff J., Payne C.C., (1973) Anal. Biochem. 55, 26.
7. Baiersdörfer M. (2019) Mol. Ther. Nucleic Acids. 15, 26.
8. Karikó, K. et al., (2008) Molecular Therapy 16, 1833.
9. Anderson, B.R. et al., (2010) Nucl. Acids Res. 17, 5884.
10. Karikó, K. et al., (2005) Immunity 23, 165.
11. Karikó, K. and Weissman, D. (2007) Curr. Opin. Drug Discov. Devel. 10, 523.
12. Andries, O. et al., (2015) J. Controlled Release 217, 337.
13. Svitkin, Y. V. et al., (2017) Nucleic Acids Res. 45 (10), 6023.

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Purchaser receives a limited non-exclusive, non-transferable right to use the Products purchased from CELLSCRIPT™ solely for its own internal laboratory research use. See the Licensing tab for more information. LIMITED LABEL LICENSE CELLSCRIPT™’s T7 mScript™ Complete Standard mRNA Production System (“Product“) is for making standard (U)-mRNA having reduced innate immunogenicity in mammalian cells. This Product and the compositions and methods and uses thereof are covered by WIPO PCT Patent Application Numbers WO2007024708A2 and WO2011071931A2 and U.S. Patent Numbers US8278036B2 and US9750824B2, and U.S. and international divisional, continuation, or other patents and patent applications derived therefrom that are owned by the University of Pennsylvania and licensed to CELLSCRIPT™.

The Min-Immune™ Gold dsRNA Removal Module of this Product effectively removes double-stranded RNA contaminants that are generated as by-products of making mRNA using a process comprising in vitro transcription (IVT) of a DNA template encoding said mRNA. This Module and the compositions and methods and uses thereof are covered by WIPO PCT Patent Application Number WO2013102203A2 and U.S. Patent Number US12059479B2 and U.S. and international divisional, continuation, or other patents and patent applications derived therefrom that are owned by CELLSCRIPT™.

By purchasing Product from CELLSCRIPT™ or an authorized distributor of CELLSCRIPT™ (herein, “Purchaser”), Purchaser receives a limited non-exclusive, non-transferable, non-sublicensable right (“Limited License”) to use the purchased Product solely for its own internal laboratory research use (the “Licensed Use”) and Purchaser agrees to cite the name of the Product and CELLSCRIPT™ as its source in any verbal or written public disclosure about said Licensed Use. The Licensed Use expressly excludes any Commercial Use comprising [A] any diagnostic, prophylactic, clinical, therapeutic or other use in humans, [B] any veterinary, livestock, agricultural use in animals, [C] making or using U-mRNA encoding transcription factors or cell reprograming factors to generate human induced pluripotent stem cells (iPSCs), hematopoietic stem or progenitor cells, neurons, immune cells or any other differentiated cells, and/or [D] manufacture, distribution, importation, exportation, or sale of any other products and/or services made using Product for which Purchaser receives compensation of any kind, it being understood that [A]–[D] are separately and/or collectively defined as a “Commercial Use” herein. No other license rights are granted, expressly or implied, to make, have made, import, export, use, reverse engineer, transfer, offer for sale, or sell Product or any other CELLSCRIPT™ product or service. By purchase of Products, Purchaser agrees not to make, have made, import, export, use, reverse engineer, transfer, offer for sale, or sell Products, components of Products, or derivatives of Products, or to provide services, information, or data obtained by the use thereof in exchange for money or other consideration. CELLSCRIPT™ provides no warranties (statutory or implied) concerning non-infringement of intellectual property rights of any other parties, and all such warranties are expressly disclaimed. This Limited License terminates automatically if Purchaser breaches any of the terms herein.

The license remains in effect if the product is being used in accordance with the license terms. The license is terminated immediately upon user’s breach of any of the license terms, such as using the product for commercial purposes without proper authorization.

If your organization is interested in using T7 mScript™ Complete Standard mRNA Production System or any other CELLSCRIPT™ products for a Commercial Use, whether in catalog or custom sizes, please contact customerservice@cellscript.com with a description of your interest and needs.

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• Generate canonical (GAUC) mRNA with greatly reduced double-stranded RNA (dsRNA) content.
• Easily remove dsRNA without the need for chromatographic methods like HPLC.
• Produce large amounts of mRNA with virtually 100% transcript capping.
• Reduce the innate immunogenic response to mRNA in cells.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200