Cambio - Excellence in Molecular Biology

Aptamers
Antibodies
Biopharma Quality Control
Buffers (custom)
Cell Culture Contamination Control
Chromosome Painting
Competent Cells
CRISPR-Cas 9
DNA and RNA (human placental)
DNA & RNA Sequencing
Extraction & Purification - DNA
Extraction & Purification - RNA
Enzymes for Molecular Biology
Gel Electrophoresis
In Vitro Toxicology
In vitro Transcription
Lab Cleaning
Lab Equipment
Lab Reagents
Oligo Synthesis
PCR
PCR diagnostics kits
qPCR and qRT-PCR
Peptide Synthesis
Protein Purification
Sampling swabs
STEM Learning
Transfection-EZ Biosystems
Transfection-Biontex
Water Contamination Testing
Search
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.

MessageMAX™T7 ARCA-Capped Message Transcription Kit

For co-transcriptional capping of IVT RNA with T7 RNA polymerase and standard cap analog. A reaction produces 60 µg RNA from 1 µg DNA in 30 minutes. Reaction products are 40% correctly capped, 40% incorrectly capped and 20% uncapped.

CellScript

Catalogue No.DescriptionPack SizePriceQty
  • Change to:
C-MMA60710MessageMAX™T7 ARCA-Capped Message Transcription Kit10 Reactions �323.15 Quantity
Add

  • Co-transcriptionally cap uridine-containing IVT RNA with Anti-Reverse Cap Analog (ARCA).
  • Optimized for maximum transcript capping and yield.
  • Produce up to 60 µg of ARCA-capped RNA from 1 µg of template in just 30 minutes.
  • ScriptGuard™ RNase Inhibitor included to protect synthesized RNA from RNase degradation.

Product is for research use only (RUO)

Product Description:

The MessageMAX™ T7 ARCA-Capped Message Transcription Kit V2 is optimized for high-yield synthesis of canonical uridine-containing Anti-Reverse Cap Analog (ARCA)-capped RNA from an in vitro transcription (IVT) reaction. A 30 minute, 20 μl reaction yields up to 60 μg of ARCA-capped RNA from 1 μg of the control template DNA. ARCA ensures correct 5′ end incorporation, resulting in RNA that translates more efficiently in vivo than standard cap analogs. ScriptGuard™ RNase Inhibitor is included in the kit to protect synthesized RNA from RNase degradation.

The ARCA / NTP PreMix contains all four ribonucleotides (canonical nucleotides GTP, ATP, UTP and CTP) and the ARCA. The PreMix ensures the optimal concentration of each NTP and ratio of ARCA to GTP (4:1), maximizing transcript capping (~80%) and yield.

MessageMAX™ ARCA-capped RNA can be further processed into mRNA for expression in cells through the use of post-transcriptional capping using CELLSCRIPT™'s ScriptCap™ 2'-O-Methyltransferase (for conversion to a Cap 1 structure) and post-transcriptional tailing using A-Plus™ Poly(A) Polymerase Tailing Kit (available separately).

For reduction of immunogenicity, combine ARCA-capped mRNA with the Min-Immune™ Gold dsRNA Removal Kit to produce virtually dsRNA-free (to <0.005% of sample [LLOQ]), ultra-low immunogenicity mRNA that is suitable for downstream applications such as cell and gene therapy research and mRNA vaccine development.

CELLSCRIPT™ also offers the INCOGNITO™ T7 ARCA 5mC- & Ψ-RNA Transcription Kit to produce low immunogenicity ARCA-capped RNA containing the modified nucleotides 5-methyl-cytidine and pseudouridine.

Materials Supplied:

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

 

 

MessageMAX™ T7 ARCA-Capped Message Transcription Kit V2
Contents (10 reactions)
Kit Component Volume
MessageMAX™ T7 Enzyme Solution 20 μl
10X MessageMAX™ T7 Transcription Buffer II 20 μl
MessageMAX™ ARCA Cap/NTP PreMix
(22.5 mM ATP, CTP & UTP; 4.5 mM GTP & 18 mM ARCA) 		80 μl
20 mM GTP 20 μl
100 mM Dithiothreitol (DTT) 20 μl
RNase-Free DNase I, 1 U/μl 10 μl
ScriptGuard™ RNase Inhibitor, 40 U/µl 10 μl
T7 Control Template DNA, 0.5 µg/µl 10 μl
RNase-Free Water 1.4 ml

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

  • A DNA template for transcription of your RNA of interest
  • Materials or kits for purification of the RNA product
  • RNase-free TE Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA)
  • Optional: TE saturated phenol/chloroform, 0.5-1 M EDTA

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

Protocols for: MessageMAX™ T7 ARCA-Capped Message Transcription Kit 

 

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to CELLSCRIPT’s product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

MessageMAX™ T7 ARCA-Capped Message Transcription Kit Protocol

Please note: all protocols are the responsibility of the product supplier

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

REFERENCES

1.Stepinski, J. et al., (2001) RNA 7, 1486.

2.Peng, Z-H et al., (2002) Org. Lett. 4, 161.

3.Jemielity, J. et al., (2003) RNA 9, 1108.

4.Warren, L. et al., (2010) Cell Stem Cell 7, 618.

5.Karikó, K. et al., (2008) Molecular Therapy 16, 1833.

6.Anderson, B.R. et al., (2010) Nucl. Acids Res. 17, 5884.

7.Karikó, K. et al., (2005) Immunity 23, 165.

8.Karikó, K. and Weissman, D. (2007) Curr. Opin. Drug Discov. Devel. 10, 523.

9.Sambrook, J. et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed.), New York, Cold Spring Harbor Laboratory Press.

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

Anti-reverse cap analogs (ARCAs) are covered by U.S. Patent No. 7,074,596, and other patents issued or pending, licensed or assigned to CELLSCRIPT, Inc. These products are accompanied by a limited non-exclusive license for the purchaser to use the purchased product(s) solely for life science research. Purchase of these products does not grant rights to: (1) offer products, components of products or any derivatives thereof for resale; or (2) to distribute or transfer the products, components of products, or any derivatives thereof to third parties. Contact CELLSCRIPT for information on licenses for uses other than life science research

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