EZ Cap™ Cy5 EGFP mRNA (5-moUTP)

EGFP mRNA with Cap 1 structure, modified by 5-moUTP and Cy5-utp, providing higher transcription efficiency and suppressing RNA-mediated innate immune activation

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Background

EZ Cap™ Cy5 EGFP mRNA (5-moUTP) will express enhanced green fluorescent protein(EGFP) once entering cells originally isolated from the jellyfish(Aequorea victoria). You can observe the green fluorescence at 509 nm. EGFP is usually used as a reporter for gene regulation and function study. It is applicable in assays for mRNA delivery, translation efficiency, cell viability and in vivo imaging etc.
EZ Cap™ Cy5 EGFP mRNA (5-moUTP) is provided at a concentration of 1mg/ml. It is co-transcriptional capped by EZ Cap™ Reagent AG (3' OMe) (Catalog No. B8178) which generates a cap 1 structure with high efficiency. Cap 1 structure is more ideal for mammalian systems and possess higher transcription efficiency than Cap 0 structure (ARCA and mCap). EZ Cap™ Reagent AG (3' OMe) obtain the ability only inserting in the proper orientation, results in forming mRNAs that can be translated twice efficiently as those initiated with EZ Cap™ Reagent AG. The addition of 5-moUTP (5-Methoxy-UTP) and poly(A) tail suppress RNA-mediated innate immune activation and increase the stability and lifetime of the mRNA in vitro and in vivo. Poly(A) tail also plays an important role in enhancing the efficiency of translation initiation. Cy5 is a synthetic red fluorescent dye with maximum excitation and emission wavelengths of 650 nm and 670 nm, respectively. Cy5 EGFP mRNA is an ideal molecule to determine mRNA delivery and localization independent of translation. Cy5-UTP and 5-moUTP is used in a ratio of 1:3 when transcribed. Substitution in this ratio results in mRNA that is easy to visualize and still can be translated in cell culture. In the rough, there is an anti-correlation between translation efficiency and Cy5-UTP substitution.
All the modifications is intention to mimics a fully processed mature mRNA. EZ Cap™ Cy5 EGFP mRNA (5-moUTP) is an ideal product to observing mRNA delivery, localization, translation and other behaviors.