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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™ N1meΨ-mRNA Production System

INCOGNITO™ T7 mScript™ N1meΨ-mRNA Production System

CellScript

Catalogue No.DescriptionPack SizePriceQty
  • Change to:
IMMY240625INCOGNITO™ T7 mScript™ N1meΨ-mRNA Production System25 rxns POA Quantity
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  • All-in-one kit to produce N1-methyl-pseudouridine-containing, fully 5’ capped and 3’ poly(A) tailed mRNA (N1meΨ-mRNA).
  • Combine the benefits of high-yield transcription with post-transcriptional capping and tailing.
  • Produce large amounts of N1meΨ-mRNA with virtually 100% transcript capping.
  • Create variable 3’ poly(A) tail lengths, from 50 to >300 A’s.
  • Lower immunogenicity in mammalian cells through incorporation of N1meΨ in synthesized mRNA.

Product is for research use only (RUO)

Product Description: 

The INCOGNITO™ T7 mScript™ N1meΨ-mRNA Production System provides all enzymes and reagents for making low immunogenicity N1-methyl-pseudouridine (N1meΨ)-containing, 5'-capped and 3'-polyadenylated mRNA. The kit includes reagents for four 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 and (4) 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. Additionally, it has been shown that N1meΨ-mRNAs are translated into protein at higher levels and induce lower innate immune responses in humans and other mammalian cells that express various RNA sensors compared to corresponding unmodified mRNAs. The INCOGNITO™ mScript™ System generates modified mRNA with virtually 100% transcript capping and user-defined poly(A) tail length. Poly(A) tail lengths can be generated much longer than is possible using a template-encoded tail, even greater than 300 A’s. This mRNA is suitable for use in transfection and microinjection experiments as well as in vitro translation systems.

For maximum reduction of immunogenicity, combine INCOGNITO™ N1meΨ-mRNA with the Min-Immune™ Gold dsRNA Removal Kit to produce virtually dsRNA-free, ultra-low immunogenicity mRNA that is suitable for 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. See the Licensing tab for more information.

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

 

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

 

Troubleshooting guide:

 

 

 

FAQs: 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.

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

Uses and Label Licenses for Specific Products

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 (the “Products”) 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

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