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In vitro Transcription

In vitro Transcription: INCOGNITO™ IVT and mRNA Production Systems

The new INCOGNITO™ kits

 

Evade cellular immune response  to your RNA with the new INCOGNITO™ kits  from Cellscript.

The new INCOGNITO™ kits *  are designed  for in vitro synthesis of RNA containing modified nucleosides such as pseudouridine and 5-methylcytidine. They assist in reducing immune response and enhancing protein translation.  A range of these kits are available to end users depending on their specific research desires and downstream applications.

Cellscript’s INCOGNITO™ contain everything needed to make N1meΨ- or Ψ-containing Cap1 mRNA, including in vitro transcription reagents plus ScriptCap™ Capping Enzyme, ScriptCap™ 2’-O-Methyltransferase, and A-Plus™ Poly(A) Polymerase.  

The INCOGNITO™ T7-FlashScribe™ N1meΨ-RNA Transcription Kit is specially formulated to enable users to obtain the maximum possible yields of N1-methyl-pseudouridine-containing RNA (N1meΨ-RNA) from an in vitro transcription (IVT) reaction. Although yield varies with the DNA template and other factors, a standard 60 minute, 20 μl reaction will yield up to 160 μg of RNA from 1 μg of the control template. These yields are made possible by the high-performance properties of the T7-FlashScribe™ enzyme. The standard reaction can be scaled up to produce milligram amounts of RNA containing the canonical nucleotides ATP, CTP, GTP and the modified nucleotide N1-methyl-pseudouridine-5'-triphosphate (N1meΨTP).

The INCOGNITO™ T7-FlashScribe™ Ψ-RNA Transcription Kit is specially formulated to maximize yields of pseudouridine-containing RNA (Ψ-RNA) from an in vitro transcription (IVT) reaction. Although yield varies with the DNA template and other factors, a standard 60 minute, 20 μl reaction will yield up to 160 μg of RNA from 1 μg of DNA template. These impressive yields are due to the high-performance properties of the T7-FlashScribe™ enzyme. The standard reaction can be scaled up to generate milligram amounts of RNA containing the canonical nucleotides ATP, CTP, GTP and the modified nucleotide pseudouridine-5'-triphosphate (ΨTP).

 

INCOGNITO™ T7 mScript™ mRNA Production Systems contain everything needed to make N1meΨ- or Ψ-containing Cap1 mRNA, including in vitro transcription reagents plus ScriptCap™ Capping Enzyme, ScriptCap™ 2’-O-Methyltransferase, and A-Plus™ Poly(A) Polymerase. 

 

The INCOGNITO™ SP6 Ψ-RNA Transcription Kit -optimized for high-yield synthesis of pseudouridine-containing RNA (Ψ-RNA) from an in vitro transcription (IVT) reaction.

The INCOGNITO™ T7 5mC- & Ψ-RNA Transcription Kit - optimized for high-yield synthesis of 5-methyl-cytidine- pseudouridine-containing RNA (5mCΨ-RNA) from an in vitro transcription (IVT) reaction.

Advantages

  • Optimized for high-yield synthesis of pseudouridine-containing IVT RNA (Ψ-RNA)
  • Produce up to 100 µg of Ψ-RNA from 1 µg of template ( depending on kit used).
  • Easily scale up reactions to produce milligram amounts of Ψ-RNA.
  • Lower immunogenicity in mammalian cells through incorporation of Ψ in synthesized mRNA.
  • Modified Ψ-mRNAs translate into higher protein levels
  • The standard reactions can be scaled up to produce milligram amounts of RNA canonical nucleotides ATP and GTP and modified pseudouridine-5'-triphosphate (ΨTP) and 5-methyl-cytidine-5'-triphosphate (5mCTP).


 

*Transcription Kits are for in vitro transcription of N1meΨ- or Ψ-containing RNA. - All kits above come with a label license to use them for internal research use under the University of Pennsylvania’s modified mRNA technologies patents.

INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System

INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System

CellScript

Catalogue No.DescriptionPack SizePriceQty
  • Change to:
IMCMY250225INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System25 rxns POA Quantity
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  • Complete solution to produce N1-methyl-pseudouridine-containing, fully
    5’ capped and 3’ poly(A) tailed mRNA (N1meΨ-mRNA) with dsRNA removal to <0.005% (LLOQ).
  • Achieve ultra-low immunogenicity in mammalian cells by synthesizing transcripts incorporating N1meΨ followed by highly efficient removal of dsRNA.
  • Produce 1.5 mg of N1meΨ-mRNA with virtually 100% transcript 5’ capping.
  • Create variable 3’ poly(A) tail lengths, from 50 to >300 A’s.

Product is for research use only (RUO) 

Product Description: 

The INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System provides all enzymes and reagents for making ultra-low immunogenicity N1-methyl-pseudouridine (N1meΨ)-containing, 5'-capped, and 3'-polyadenylated mRNA with ultra-low double-stranded RNA (dsRNA) content. The kit includes reagents for five workflow modules, (1) high-yield in vitro transcription of linear double-stranded DNA templates using the T7 mScript™ Enzyme Solution, the canonical nucleotides ATP, CTP, GTP and the modified nucleotide N1-methyl-pseudouridine-5'-triphosphate (N1meΨTP), (2) 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, (3) A-Plus™ Poly(A) Polymerase for adding a 3'-poly(A) tail, (4) post-transcriptional enzymatic removal of dsRNA content using the Min-Immune™ Gold dsRNA Removal system and (5) 5 M NH4OAc as a convenient RNA/mRNA purification method.

Post-transfection, 5’ capped and 3’ poly(A) tailed mRNA has increased stability and translation efficiency in most eukaryotic cell lines. The INCOGNITO™ mScript™ System generates modified mRNA with virtually 100% transcript capping and user-defined poly(A) tail length. The A-Plus™ Poly(A) Polymerase protocol provides a wide range of poly(A) tail lengths from 150–300 bases. The included Min-Immune™ Gold dsRNA Removal module can reduce dsRNA to <0.005% (LLOQ) of the sample, lower than what can be achieved using engineered mutant T7 RNA polymerases. Ultra-low immunogenicity INCOGNITO™ Complete N1meΨ-mRNA is suitable for use in downstream applications such as cell and gene therapy research and mRNA vaccine development.

Uses and Label Licenses for Specific Products:

Purchaser receives a limited non-exclusive, non-transferable right to use the Products purchased from CELLSCRIPT™ solely for its own internal laboratory research use.

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

Terms & Conditions: The Product listed is currently offered by CELLSCRIPT™ for research use only under the defined Terms and Conditions

Troubleshooting Guide:

 

 

For answers to Frequently Asked Questions about our products, visit the FAQ library.

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

Manual

SDS

These are both links to the manufacturer's page

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

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

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

LIMITED LABEL LICENSE

CELLSCRIPT™’s INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System (“Product”) is for making N1meΨ-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 N1meΨ-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 INCOGNITO™ T7 mScript™ Complete N1meΨ-mRNA Production System or any other in 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.

Research Use Label License

INCOGNITO™ kits or other products or services comprising N1-methyl-pseudouridine, pseudouridine, 5-methyl-cytidine and/or other modified nucleotides for use in making RNA or mRNA that are covered by technologies and IP disclosed in U.S. Patent Nos. 8,278,036 and 9,750,824 and PCT Publ. Nos. WO 2007/024708 and WO/2011/071931 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 (“Products”) are sold for research use only.

By purchasing these Products from CELLSCRIPT or an authorized distributor of CELLSCRIPT, Purchaser receives a limited non-exclusive, non-transferable right to use the Products purchased from CELLSCRIPT solely for its own internal laboratory research use (the “Product Use”). Such Product Use expressly excludes any Commercial Use comprising [A] clinical or other use in humans, [B] veterinary, livestock, agricultural or commercial use in animals, and/or [C] manufacture, distribution, importation, exportation, or sale of any products and/or services made using Products, it being understood that [A]–[C] are separately and/or collectively defined as a “Commercial Use” herein. By purchase of Products, Purchaser agrees not to make, import, use, transfer, offer for sale, or sell Products, components of Products, or derivatives of Products, including modified nucleoside-containing RNA or cells made by use 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. Please contact CELLSCRIPT to inquire about a license for a Commercial Use. The license remains in effect as long as the product is being used in accordance with the license terms.

The license is terminated if the user breaches any of the license terms, such as using the product for commercial purposes without proper authorization.

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