mRNA encoding Enhanced green fluorescent protein
eGFP mRNA encodes Enhanced Green Fluorescent Protein (eGFP), the most widely used fluorescent reporter in cell biology and molecular research. eGFP is a humanized variant of wild-type GFP originally isolated from the jellyfish Aequorea victoria, engineered to overcome the limitations of the native protein. Two key point mutations, F64L and S65T, dramatically improve performance in mammalian systems: F64L enhances chromophore maturation at 37°C, while S65T simplifies the excitation spectrum to a single peak at ~488 nm and increases brightness approximately 35-fold over wild-type GFP. The coding sequence was further optimized with over 190 silent nucleotide substitutions to reflect preferred human codon usage, yielding an additional ~4-fold increase in mammalian expression. The resulting protein fluoresces bright green (excitation 488 nm, emission 507 nm), requires no exogenous cofactors, and is compatible with standard fluorescein filter. The mRNA construct includes a 5' Cap1 structure and poly(A) tail to support efficient translation in mammalian cell-free and cellular expression systems.
Applications:
1) Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W., & Prasher, D. C. (1994). Green fluorescent protein as a marker for gene expression. Science, 263(5148), 802–805
2) Cormack, B. P., Valdivia, R. H., & Falkow, S. (1996). FACS-optimized mutants of the green fluorescent protein (GFP). Gene, 173(1), 33–38
3) Yang, T. T., Cheng, L., & Kain, S. R. (1996). Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein. Nucleic Acids Research, 24(22), 4592–4593
4) Tsien, R. Y. (1998). The green fluorescent protein. Annual Review of Biochemistry, 67, 509–544
5) Zeng, C., Hou, X., Yan, J., Zhang, C., Li, W., Zhao, W., Du, S., & Dong, Y. (2020). Leveraging mRNA sequences to express SARS-CoV-2 antigens in vivo. Cold Spring Harbor Laboratory
Cisterna's Off-the-shelf are validated to be purchased with a quick quote generated by the system. This means you go through a series of quick steps as below to complete your purchase. Step 1: If you already know the RNA you want, select the RNA you desire on the main page by clicking the 'Quality', 'Amount', and 'Quantity' on the main page and clicking on 'Add to Cart' button. If you want to read more about the individual RNA, click on the category of your RNA and select the RNA you desire by directly clicking on its name. This will take you to the individual RNA page, where you can learn more about the RNA, its usage in research with top examples for each of the use cases under the 'References' tab. Step 2: Once you are ready to order, just select the 'Quality', 'Amount', and 'Quantity', then click on 'Add to Cart' button. Step 3: After all the items have been added to the Cart, Click on 'Get a quote' button. Step 4: Fill in all of the information such as name, your professional email, and academic or business address. Note that non-professional emails and home addresses will not be considered. Step 5: Once the information is filled in, you may download the quote as PDF and save it for your reference. Step 6: You can then click on 'Continue to checkout' button which will take you to the final 'Checkout' page. Step 7: You can provide an optional Purchase Order (PO) number here as some institutions may require this. If everything looks ok, then click on 'Place an order'. Thats it! You are done. We will take it from here. We will make your RNA and ship in the fastest possible time. Invoices will be sent electronically to the email after confirmation of delivery. All invoices have a Net30 agreement.
PRIME is the standard quality RNA made by Cisterna. All RNAs including mRNAs are made with PRIME quality. The PRIME platform generates very low impurities such as dsRNA, short-truncated products, abortive transcripts and DNA:RNA hybrids. The result is a highly pure RNA that is functional and can be used for most downstream applications demanding high-purity such as X-ray crystallography, Cryo-EM, tRNA aminoacylation and to generate maximum amount of functional proteins.
Both PRIME and SPECC reflect Cisterna's commitment to high-quality RNA, but they are designed for different needs and applications. PRIME is Cisterna's standard production system and is applicable to all RNA types. It delivers very high purity RNA suitable for a wide range of demanding applications, from structural studies to functional protein expression. SPECC (SPEcific Capture and Cleavage) is Cisterna's flagship system and is currently applicable to mRNA only. It takes purity a step further by selectively isolating full-length mRNA transcripts, approaching HPLC-level purity. SPECC is the ideal choice when the highest possible mRNA quality is required, particularly for quantitative studies, high-dose applications, and therapeutic development. In terms of cost, SPECC is approximately 5-10x the price of PRIME, reflecting the advanced purification processes involved. While affordably priced, PRIME still delivers very high RNA quality, well above what you might expect at this price point.