Enzymatic Protein Biopolymers as a Tool to Synthetize Eukaryotic Messenger Ribonucleic Acid (mRNA) with Uses in Vaccination, Immunotherapy and Nanotechnology

Abstract

Multi-subunit enzymes are protein biopolymers that are involved in many cellular processes. The enzyme that carries out the process of transcription of mRNAs is RNA polymerase II (RNAPII), which is a multi-subunit enzyme in eukaryotes. This protein biopolymer starts the transcription from specific sites and is positioned by transcription factors, which form a preinitiation complex (PIC) on gene promoters. To recognize and position the RNAPII and the transcription factors on the gene promoters are needed specific DNA sequences in the gene promoters, which are named promoter elements. Those gene promoter elements can vary and therefore several kinds of promoters exist, however, it appears that all promoters can use a similar pathway for PIC formation. Those pathways are discussed in this review. The in vitro transcribed mRNA can be used as vaccines to fight infectious diseases, e.g., in immunotherapy against cancer and in nanotechnology to deliver mRNA for a missing protein into the cell. We have outlined a procedure to produce an mRNA vaccine against the SARS-CoV-2 virus, which is the causing agent of the big pandemic, COVID-19, affecting human beings all over the world. The potential advantages of using eukaryotic RNAPII to synthetize large transcripts are outlined and discussed. In addition, we suggest a method to cap the mRNA at the 5′ terminus by using enzymes, which might be more effective than cap analogs. Finally, we suggest the construction of a future multi-talented RNAPII, which would be able to synthetize large mRNA and cap them in the test tube.

Smart Polymeric Nanoparticles in Cancer Immunotherapy

“…These biopolymers are biodegradable and biocompatible. These natural biopolymers can be combined with nanotechnology to provide multifunctional nanodrug carriers for cancer immunotherapy [ 190 ]. The nanoformulation reduces the cytotoxicity of the payload or coating carrier and prevents the degradation of the drug.…”

mRNA vaccine: a potential therapeutic strategy

“…The m(7(LNA))G[5′]ppp[5′]G 3 cap analogue increases the translational efficacy [51]. Apart from various modification forms of 5′ cap, like 7-methylguanosine [52], capping the mRNA at the 5′ terminus with enzymes is more effective than various forms of cap analogs [53,54]. The poly(A) tail is another mRNA-stabilizing element.…”

Recent Advances and Innovations in the Preparation and Purification of In Vitro-Transcribed-mRNA-Based Molecules

“…In contrast, co-transcriptional capping using cap analogues does not require a second enzymatic reaction and only needs a nucleic acid precipitation or purification process in the IVT process [ 37 ]. Therefore, co-transcriptional capping can be a more efficient process, presumably leading to a lower cost of time and goods in a research laboratory [ 38 , 39 ], but the first-generation cap analogue resulted in Cap 0 structures with low capping efficiency (60–80%) [ 40 ]. The technology for a one-pot synthesis of capped IVT mRNA is improving with the development of new cap analogues.…”