Tools: mRNA Standards Application Cases

Mondo Technology Updated on 2024-02-05

Background:

The successful development of mRNA vaccines for the novel coronavirus has brought unprecedented attention to mRNA technology. As a general-purpose technology, mRNA technology is expected to be widely used in many fields such as infectious disease vaccines, tumor vaccines, immunity, protein substitution, and sexual antibodies, leading pharmaceutical research and development into a new era.

Tool mRNA plays a very important role in the study of nucleic acid drug delivery systems, the efficacy study of protein expression, and the development of mRNA drugs.

What is Tool mRNA?

The tool mRNA (mRNA standard) is the standard messenger RNA of the finished product (similar to the plasmid commonly used in molecular experiments). The mRNA standard does not need to be processed after purchase and can be directly applied to the corresponding experiment.

ABS has a range of linear, circular tool mRNAs. Among them, the linear mRNA adopts the CAP1 cap structure synthesized by chemical chemistry, so that the linear mRNA has a high proportion of natural cap structure, and there are 5 modified bases (N1-methylpseudo-UTP, pseudo-UTP, 5-methoxy-utp, 5-methyl-CTP, N6-methyl-ATP), and the codon, UTR, and polya structures suitable for mammalian cells are optimized. Circular mRNA is generated by in vitro transcription and circularized, and then isolated and purified to obtain a finished product.

Tool mRNA Classification:

1. Reporter genomics

Reporter mRNAs are commonly used in cell transfection, translation efficiency, drug delivery, in vivo imaging, and more. Among them, Ebison EGFP mRNA is one of the most commonly used tracer protein mRNA, with strong fluorescence stability, used to express enhanced green fluorescent protein (isolated from aequorex), with an emission wavelength of 509nm, which can be detected by fluorescence microscopy or flow cytometry, and is an ideal tool for observing mRNA delivery and translation.

Figure of EGFP mRNA Agarose electrophoresis bands.

Figure Expression effect of HEK293 cells after transfection with EGFP mRNA for 24 hours.

ABS also has other reporter gene tools such as TDTOMATO mRNA, YFP mRNA, mBanana mRNA and other fluorescent protein mRNA, which can be used for control validation experiments, please feel free to contact us if you need it.

2. Gene editing

CRISPR Cas9 is an advanced gene editing system that is widely used in a wide range of diseases. Cas9 mRNA transiently expresses Cas9 protein within cells, reducing the risk of immune response and off-target.

3. Immune antigens

Immunoantigen-like mRNA is an ideal research tool for tumor immunity**. Ovalbumin (OVA) is a member of the filamentous protein superfamily and is the main glycoprotein found in egg whites. It is a commonly used antigen for immunological and biochemical research and is an established airway hyperreactivity model allergen that activates cellular and humoral immunity.

4. Protein substitution

EPO mRNA is used to express human erythropoietin. We produced EPO mRNA using (ABS CaptM) transcription in vitro, and the chemically synthesized Cap1 structure gives it a high proportion of native mRNA cap structure, resulting in efficient expression.

toolsmrnaProduct Features:

toolsmrnaBenefits:

1. CAP1 cap structure, improve mRNA stability and translation efficiency, more suitable for mammals;

A variety of modifications can be selected, and the addition of modified bases can improve translation efficiency and reduce immunogenicity;

3. Appropriate polya tail structure to ensure mRNA stability and translation efficiency;

4. High integrity, strict quality control, convenient for research and application.

User experiment case 1:

The mCherry mRNA was transfected into HEK293 cells with nucleic acid transfection kit (ABS60259) for 24 h and 48 h, and the red fluorescence observed under fluorescence microscopy was stable and the signal was strong. (For specific experimental operations, please refer to the experimental operation steps of nucleic acid transfection kit (ABS60259)).

Figure Fluorescence after 24 h of transfection of mCherry mRNA.

Figure Fluorescence after 48 h of transfection of mcherry mRNA.

User experiment examples

The mRNA transfection reagent (ABS60339) was used to transfect the thread *** mRNA-egfp mRNA in different types of cells, and the cells were treated for 24 hours and photographed by fluorescence microscope, and all kinds of cells showed strong green fluorescence.

Figure: Transfection of EGFP mRNA into different cell types.

User experiment examples

Linear luciferase mRNA was injected into live mice, the mouse weighed 150ug, and each needle was injected with 20ul, a total of 3 injections, and in vivo imaging was performed after 3 days.

Figure Luciferase mRNA injected into mRA in vivo after mouse imaging.

faq:

toolsmRNA preservationWhat are the conditions?

A: It needs to be stored below -40°C, -80 refrigerator or liquid nitrogen storage, the products provided by Absin are freshly produced, in order to achieve the best use effect, please use within 2 months after receiving the goods. If you accidentally store the mRNA at -20 for a short period of time (e.g., 1 day), you can run the mRNA on agarose gel electrophoresis to see if it degrades before the experiment.

Tool mrnaWhat is the status?

A: Our tool mRNA is in a liquid state, and the general solvent is generally 1mm sodium citrate salt solution, with a pH of 64。

Tools mRNAHow long is the delivery time?

A: Our company's tools mRNA are all freshly produced products, and we will arrange production and delivery immediately after receiving the order. The general delivery time is 1-2 weeks.

4. What is the purity of the tool mRNA?

A: Linear mRNA can reach 90% purity; If the circular mRNA is 3-5K in length, it can reach scientific grade purity, usually 85% purity. The longer the circular mRNA sequence, the lower the loop closure efficiency and the more difficult it is to purify.

5. Tool MrnaWhat is the recommended cap structure?

a:cap1 ag。The capping rate is as high as 95%.

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