Nucleotide Modifications of mRNA

With the development of in vitro transcription (IVT) methods, messenger ribonucleotide (mRNA) synthesis has emerged as an emerging class of gene therapy. To support mRNA research and therapeutic applications, BOC Sciences offers nucleotide-modified mRNA with industry-leading project completion times, a wide range of deliverables and best-in-class quality.

Introduction

mRNA Profile

Messenger RNA (mRNA) is a single-stranded ribonucleic acid, transcribed from a single piece of DNA, carrying the information encoded for protein synthesis, further transcription and processing into a functional protein. It is a temporary copy, so it has the characteristics of few copies, short lifespan, and few modifications.

Fig. 1 Structure of eukaryotic mRNA

Researchers have investigated many mRNA modification strategies. One of the more important discoveries is nucleotide modification, and mRNA containing various modified nucleotides can effectively reduce the immunogenicity of mRNA and inhibit innate immune activation, making mRNA a powerful tool for regenerative medicine, disease treatment and cell reprogramming.

Modified Nucleotides

The autoimmunogenicity of in vitro transcribed mRNAs is a problem that cannot be ignored, and exogenous RNA can be regarded as a signal of virus infection. Nucleotide chemical modification strategies can reduce immunogenicity without affecting its translational properties. For example, natural adenosine is replaced with N1-methyl-adenosine (m1A) or N6-methyl-adenosine (m6A); natural cytidine is replaced with 5-methyl-cytidine (m5C); natural uridine is replaced with 5-methoxyuridine (5moU), N1-methyl-pseudouridine (m1Ψ), pseudouridine (Ψ), etc. Among them, N1-methyl-pseudouridine (m1Ψ), 5-methoxyuridine (5moU), and pseudouridine (Ψ) have attracted much attention because both in vivo and in vitro experiments have shown that they are effective in reducing the immunogenicity of mRNA while significantly improving its translation efficiency.

• Pseudouridine

Pseudouridine is derived from uridine by a base isomerization reaction in which the nucleobase is rotated 180° around the N3-C6 axis, resulting in a change in the nucleobase-sugar bond and the resulting C-C bond allowing the nucleobase to rotate more freely. Pseudouridine can base pair with adenosine in the same way as uridine, but pseudouridine can alter RNA structure by improving base pairing, base stacking and backbone stability.

• N1-methyl-pseudouridine

N1-methyl-pseudouridine is a methyl-pseudouridine, an N1-modified pseudouridine derivative, a natural modification found in 18S rRNA and tRNA in many organisms. N1-methyl-pseudouridine has a methyl group at the N1 position, thereby eliminating the additional hydrogen bond donor.

• N6-Methyl-ATP

m6A is the most abundant internal modification of mRNA and long-stranded non-coding RNA in most eukaryotes. In 2012, scientists' studies showed that m6A modifications are associated with mRNA stability, splicing processing, and translation.

• 5-Methyl-CTP

m5C has been found in mRNA, rRNA and tRNA from a variety of representative organisms. As reversible epigenetic modifications, C modification of m5RNA affects the fate of modified RNA molecules, including promotion of mRNA stability, splicing and nucleoplasmic transport; viral protein expression; DNA damage repair; mRNA stability; cellular tolerance, proliferation and migration; and stem cell development, differentiation and reprogramming.

• 5-Methoxy-UTP

5-methoxyuridine is a rare nucleoside. The addition of 5-methoxyuridine to RNA (mRNA) reduces the immunogenicity of the resulting mRNA.

• Cyanine 5-UTP

Cyanine 5-UTP (Cy5-UTP) can be used as a substrate for T7 RNA polymerase instead of UTP to produce labeled probes by in vitro transcription.

BOC Sciences Provides the Following Modified Nucleotides

Advantages

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

Inquiry and Order

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