Cambio - Excellence in Molecular Biology

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.   - INCOGNITO™ T7-FlashScribe™

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.   - INCOGNITO™ T7-FlashScribe

 

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
C-MMA60710MessageMAX™T7 ARCA-Capped Message Transcription Kit10 Reactions £276.00 Quantity Add to Order

Description

Make mRNA with an ARCA Cap for increased translation efficiency 

The MessageMAX T7 ARCA-Capped Message Transcription Kit uses the Anti-reverse cap analog (ARCA) dinucleotide to initiate the transcription of capped RNA molecules. Unlike standard cap analogs, the ARCA cap cannot be incorporated in the reverse orientation, which ensures that all of the capped RNA will be active in the translation process. 

The MessageMAX T7 ARCA-Capped Message Transcription Kit provides a higher yield, up to 60 µg, than other commercial ARCA capping kits. The fast, 30 minute protocol offers significant time savings as well. 

If the capped RNA is to be used for translation studies, translational efficiencies can be much higher if the mRNA is also polyadenylated, especially if the mRNA is intended for use in vivo following transfection or microinjection. RNA can be easily and efficiently polyadenylated using the A-Plus™ Poly(A) Polymerase Tailing Kit. 

Reaction Definition: The MessageMAX™ T7 ARCA-Capped Message Transcription Kit yields up to 60 µg of RNA having an ARCA cap per µg of a 1.4-Kb Control Template in a 20-µl reaction in 30 minutes. Results may vary due to differences in templates. 

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 EPICENTRE. 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 EPICENTRE 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

Protocols

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

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

Notes

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

Applications & Benefits

Benefits

    • The ARCA cap analog can only be incorporated in the proper orientation, ensuring optimal translation efficiency form all capped transcripts
    • MessageMAX T7 reactions require only 30-minutes to produce up to 60 µg of RNA
    • Up to 80% of the RNA produced is capped
    • The ARCA Cap/NTP PreMix in the kit contains optimal levels of ARCA Cap and NTPs for typical mRNA lengths. A tube of GTP is included if exceptionally long transcripts are desired

 

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

Related products

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