>mRNA tool enzymes
In 1990, Wolff et al. Kicked off the application of mRNA in the clinical field. mRNA technology is to synthesize mRNA through in vitro transcription technology, then transport mRNA into human body by using appropriate delivery system, and translate mRNA into target protein by relying on cell's own translation system, so as to achieve the purpose of clinical treatment.
In the process of mRNA synthesis, the 5 'end of mRNA will be capped by using the vaccine capping system, the 3' end will be tailed by using poly (a) polymerase, and the highly active T7 RNA polymerase and mutant RNA monomer will be selected to improve the translation efficiency and stability, inhibite exonuclease degradation and deadenylation of mRNA.
For the series of processes of mRNA synthesis and modification, ACROBiosystems selects E.coli Expression System to develop tool enzyme products with high purity, high activity, low endotoxin, no heavy metal and host enzyme residues, including vaccinia capping system, poly (a) polymerase, T7 RNA polymerase, cap-2 '- O-methyltransferase, Pyrophosphatase, RNase inhibitor, DNase I, etc. And GMP series can be customized to meet the needs of scientific research and industrial customers.
|Vaccinia Capping System|
Add 7-methylguanosine cap structure (m7gppp, cap 0) to the 5 'end of RNA and significantly improve the stability and translation ability of mRNA
Use S-adenosylmethionine (SAM) as methyl donor, add a methyl group to the 2 '- O of the first nucleotide of the cap structure at the 5' end of RNA, and cap 0 was methylated to cap 1
Catalyze the addition of poly (a) tail at the 3 'end of RNA, has high tail addition efficiency
|T7 RNA Polymerase|
Use single or double stranded DNA containing T7 promoter sequence as template and NTP as substrate, RNA complementary to single stranded DNA downstream of promoter was synthesized
PPase can hydrolyze the inorganic pyrophosphate generated with the reaction to avoid its inhibition on the reaction system when used in nucleic acid amplification experiment
|RNase Inhibitor||Combine with RNase A, B or C in a non competitive manner 1:1, so as to inhibit the activities of three enzymes and protect RNA from degradation|
|DNase I||Any site on any strand of double stranded DNA can be randomly identified and cut in the presence of Mg2 +|
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