mRNA Reengineering Technology for Protein Production Enhancement
Creative Biolabs has a professional protein expression team with all the protein expression systems available on the market including bacteria, fungi, insects, mammals, protozoa, and plant expression systems. In order to further reduce production costs and increase protein yield, we have developed a series of effective upstream technologies to meet the growing demand for protein production.
Background
Increasing the expression level of proteins to enhance protein production has always been an important requirement for the development of protein drugs and vaccines. In general, in addition to the selection of different expression systems, attempts to induce different concentrations of IPTG, media composition, induction temperature, induction time, and codon optimization are also commonly employed strategies. However, excessive concentrations of IPTG are toxic to cells, and nutrient-rich media means higher costs. In large-scale production, besides the need to fine-tune protein expression conditions to meet specific requirements, the conditions for protein expression are generally fixed. Therefore, the design of upstream genetic constructs is another important means to increase the expression of recombinant proteins.
mRNA Reengineering Technology
The 5' cap structure of the mRNA or the internal ribosome entry site (IRES) initiates the translation of eukaryotes by recruiting 40S ribosomal subunits and other translational components. A widely accepted view is the scanning mechanism, which considers the 40S subunit to scan from the 5' end to 3’ end through the 5' leading sequence, before encountering the first AUG codon of the nucleotide. However, a recent view is that an accessible AUG codon to which the 40S subunit can move is not necessarily the first AUG codon in the mRNA. The upstream AUG codon can inhibit translation initiation by transferring the ribosome from the true initiation codon. The scanning mechanism does not take into account the effect of the potential initiation codon in the coding sequence on protein synthesis. In addition, microRNA (miRNA)-mediated down-regulation can also have a negative impact on protein translation. It can base pair with mRNA and thus reduce the stability of the mRNA and the stability of the nascent peptide chain.
Based on the above findings, we have developed a technology for reengineering mRNA to increase the yield of the protein of interest. Collectively, this technology achieves the goal of enhancing protein production through 2 aspects, one is to reduce ribosome transfer by AUG or a non-canonical initiation codon in the coding sequence, and the other is to eliminate miRNA binding sites in the coding sequence to avoid miRNA mediated down-regulation.
As a CRO/CDMO with more than 10 years of vaccine development, Creative Biolabs is not only equipped with complete technical platforms and production facilities, but also has a team of skilled and experienced researchers. Although protein expression has become a relatively mature technology in the field of biomedicine, the demand for increased production and cost compression still exists. In this regard, we have introduced an mRNA reengineering technology that can effectively achieve this goal.
All of our products can only be used for research purposes. These vaccine ingredients CANNOT be used directly on humans or animals.
