mRNA 3'-end Processing

With the emergence of mRNA vaccines driven by the new crown epidemic, mRNA holds great promise in addressing unmet therapeutic needs. BOC Sciences offers mRNA solutions to accelerate your research.

Introduction

mRNA Structure and Function

The mature mRNA structure consists of five main parts, starting from the 5'-end, the 5'-end cap region, the 5' UTR, the open reading frame encoding the antigen, the 3' UTR and the Poly(A) tail, and modification of these structures can effectively improve the stability and translation efficiency of mRNA in the cell. The 5'-end cap structure and the Poly(A)tail play a more important role in the function of mRNA.

Fig. 1 Mature mRNA in eukaryotes

Poly-adenylation

It is the covalent linking of polyadenylic acid to mRNA molecules. During protein biosynthesis, this is part of the way in which mature mRNAs are produced ready for translation. In eukaryotes, poly-adenylation is a mechanism whereby mRNA molecules are interrupted at their 3'-ends.

Poly(A) Tail

The tail is not added to the 3'-end of the transcription termination, but at the 3'-end of the transcription product. A specific enzyme recognizes the tailing of 13-20 bases upstream of the cleavage site, recognizes the signal AAUAAA and the conserved sequence GUGUGUG downstream of the cleavage site, cuts off a section downstream of the cleavage site, and then adding the Poly(A) tail, catalysed by Poly(A) polymerase. If this recognition signal is mutated, both excision and polyadenylation are significantly reduced.

Most eukaryotic mRNAs have a Poly(A) tail consisting of 100-200 A's at the 3'-end. The Poly(A) tail is not encoded by DNA, but the post-transcriptional pre-mRNA takes ATP as a precursor, and is catalyzed by RNA terminal adenylyltransferase, namely Ploy(A) polymerase, to polymerize to the 3' end.

The function of the Poly(A) tail of mRNA is

• It is a key factor in determining the lifespan of mRNA molecules
• May facilitate mRNA transport from the nucleus to the cytoplasm
• Avoid ribozyme degradation in cells, stabilize mRNA and promote protein translation. The length of the Poly(A) tail is proportional to the translation efficiency
• Acts as a recognition signal for the ribosome (when mRNA is bound to the 5'-end, this role enables the ribosome to determine whether the RNA remains intact before it consumes any energy and precursors, since tailing allows a simple of purified mRNA)

mRNA Tailing Method

• Modification by using recombinant Poly(A) polymerase, but the length of the prepared Poly(A) tail is different, which will affect the translation efficiency. This method is affected by temperature and time, RNA degradation is fast, and the ratio of RNA to ATP needs to be calculated.
• By designing a sequence encoding a fixed length in the template and directly transcribing it in vitro, this method is simple to operate and can determine the length of the Poly(A) tail.

Service Details for mRNA 3'-end Processing

Advantages

• Fast project completion
• Easy project management
• Strict quality control of mRNA
• Reliable consulting and support

Inquiry and Order

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