Overview

Efficiently generate mRNAs

When you want instant expression after transfection into cells, deliver your gene-of-interest as an mRNA made with SBI’s mRNAExpress™ mRNA Synthesis Kit. Designed to generate in vitro transcripts for transfection of mammalian cells, micro-injection into oocytes, in vitro translation, and other related applications, this high-yield kit can produce 20-40 µg of high-quality mRNAs in one standard reaction, and comes with a number of performance-enhancing features:

• Leverages the robust T7 RNA Polymerase
• Incorporates an anti-reverse cap analog (ARCA) into the transcript to increase translation efficiency
• Reduces host cell immune response and enhances stability by incorporating modified nucleotides (5-Methylcytidine-5′-Triphosphate and Pseudouridine-5′-Triphosphate) and a poly-A tail
• Degrades the DNA template after RNA synthesis with the included DNase
• Removes the 5′ triphosphates at the end of the RNA with the included phosphatase to further reduce innate immune responses in mammalian cells
• Employs a robust clean-up spin column system that delivers high yields of mRNAs that are ready for most downstream applications

The Kit includes the following reagents that are critical in vitro transcription:

• pMRNA^XP mRNAExpress Vector so you can clone and then transcribe your cDNA-of-interest
• Sequencing primers to confirm the identity of the cDNA insert cloned into the pMRNA^XP Vector
• Tail PCR primer mix to add a poly-A tail to the DNA template
• In vitro transcription reagents—T7 RNA polymerase, in vitro transcription buffer, NTP/Cap mix, RNase-free DNase, phosphatase buffer and RNase-free phosphatase
• Clean-up spin column system, including binding, wash, and elution buffers, collection tubes, and collection tubes with filter cartridges

How It Works

The mRNAExpress mRNA Synthesis Kit workflow

The mRNAExpress mRNA Synthesis Kit has a straightforward workflow:

1. Clone the cDNA of your gene-of-interest in the pMRNA^XP mRNAExpress Vector (included in the Kit, but also available separately, # MR000PA-1)
2. Tail-PCR amplify the in vitro transcription template from the pMRNA^XP Vector with your insert cloned in. The tail PCR reaction adds a poly-A tail to the DNA template. Purify the template using a spin column.
3. Perform the in vitro transcription reaction (uses the ARCA and modified nucleotides)
4. Remove the template DNA and the terminal 5′ triphosphates with the included DNase and phosphatase.
5. Prepare the mRNA transcripts for transfection with the included clean-up spin column system.
6. Measure the mRNA concentration and you are ready for transfection.

Supporting Data

Get instant expression when you deliver mRNAs made with the mRNAExpress Vector mRNA Synthesis Kit

Figure 1. Strong GFP and RFP expression after transfection with mRNAs made with the mRNAExpress Vector mRNA Synthesis Kit. GFP and RFP mRNAs generated using the mRNAExpress mRNA Synthesis Kit were transfected into human foreskin fibroblasts (HFFs) using 1 µg of each mRNA per 24-well plate of cells.

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