The mScript™ mRNA Production System provides a quick and easy way to build a superior capped and polyadenylated mRNA transcript. Based upon a high-yield T7 transcription system, the all-inclusive kit contains the reagents needed to produce high yields of 100% capped mRNA; all you need is the template DNA.

The mScript™ mRNA Production System differs from co-transcriptional mRNA production methods by decoupling the capping and transcription processes. Instead of using a costly and inefficiently incorporated nucleotide cap analog, the mScript™ mRNA Production System uses the Vaccinia virus-derived capping enzyme to add the cap, ensuring 100% capping and 100% proper cap orientation. In addition, the mScript™ system also contains the Vaccinia 2'-O-Methyltransferase enzyme, allowing the natural, translation-boosting Cap 1 structure to be built.

After capping, poly(A) tails of the desired length can be added with the included mScript™ poly(A) polymerase enzyme. Each reaction yields approximately 60 ug of fully capped and polyadenylated mRNA ready for use in transfections, microinjections, or in vitro translation systems.

Figure 1  mScript™ mRNA produces more luciferase activity than co-transcriptionally capped mRNA. Equal amounts of mScript™-produced Cap 0-capped RNA, Cap 1-capped RNA, and Cap 0 mRNA produced co-transcriptionally with standard cap analog (Std CA) or anti-reverse cap analog (ARCA) were transfected into HeLa cells, NIH3T3 cells, and BDCM cells. In vivo protein expression was measured by determining the average RLU per microgram of total protein for each cell type 24 hours after transfection. The RLU data is normalized to the activity obtained from the mScript™ Cap 0-capped RNA (100%).

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Protocols for: mScript™ mRNA Production System

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T7 mScript™ Standard mRNA Production System

(catalogue number MSC11625 / MSC11610)

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

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References:

1. Nicholson, B. L., et al. (2010) Tombusvirus recruitment of host translational machinery via the 3' UTR, RNA 16 , 1402-1419.
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3. Palavicini, J. P., et al. (2009) An extra double-stranded RNA binding domain confers high activity to a squid RNA editing enzyme, RNA 15 , 1208-1218.

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Benefits

• 100% capping - Virtually all transcripts will be capped; co-transcriptional systems typically cap less than 80%.
• 100% proper cap orientation - Traditional co-transcriptional systems use an expensive cap analog which can be incorporated in the reverse orientation; these transcripts are not translated by the cell, reducing translation efficiency.
• Cap 1 structure - mScript is the only mRNA production system that builds this translation boosting structure; Cap 1 mRNA exhibits up to 50% better translation in vivo.
• Yield - The decoupling of the transcription and capping processes allows for much higher yields than most co-transcriptional system; up to 60ug per reaction.
• Your time - Higher yields, more capping, and the better in vivo translation, all in the same amount of time as the competition.

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