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CD BioSciences' multi-molecular discovery platform based on mRNA display technology will promote the development and application of high-throughput in vitro screening technology, and is committed to becoming a leader in the development of multifunctional molecules in cell-free systems. The platform revolves around the design of special sequences, the construction of large-capacity libraries, unlimited screening and structural modification from parts to wholes, enabling it to provide one-stop molecular discovery services for global customers.
Currently, in vitro directed evolution has been widely used to screen nucleic acid molecules with improved binding or catalytic properties. Since proteins play an important role in biology and have been widely used in diagnostics, pharmaceuticals, and industry, scholars have shown great interest in the development of in vitro screening and directed evolution methods for proteins. mRNA display is a technique for creating ultra-diverse peptide and protein libraries (trillions of independent sequences) for in vitro selection and directed evolution. Using mRNA display, it is possible to select and evolve polypeptides with distinct functional properties, such as binding proteins, RNAs, post-translational modifications, and small molecules.
Fig. 1 Directed evolution process. (Kamalinia G, et al., 2021)
mRNA display technology, also known as mRNA-protein fusion display technology, is an in vitro polypeptide/protein screening technology that fuses genotype (mRNA) and phenotype (protein), and can be used for the discovery and interaction analysis of biomolecular ligands. The mRNA display technology mainly has 6 major steps. The core element of mRNA display technology is puromycin, and the core step is to form mRNA-protein complexes through puromycin.
The puromycin is connected to the 3' end of the mRNA through an oligonucleotide linker, and the mRNA with puromycin is used as a template through the in vitro translation of ribosomes to carry out the coding sequence and translate it into a nascent peptide chain. At this time, puromycin will compete with the aminoacylated tRNA for inhibition, and the former enters the A site of the ribosome, and will form a stable bond with the C-terminal amino acid of the nascent peptide chain through the structural O-methyltyrosine. Amide bonds are used to achieve covalent linkage, generate mRNA-peptide complexes, and perform affinity purification to remove ribosomes and other components in the reaction solution.
The mRNA display technology belongs to the in vitro display technology. Compared with the in vivo display technology including the phage display technology, it has many advantages, mainly reflected in the diversity and properties of the library, the screening process and the experimental cycle.
| Library Diversity | Library Properties | Reaction Conditions | Screening Cycle |
|---|---|---|---|
| The expression and screening of the mRNA display technology are performed in vitro without relying on cells, so the steps of transformation and cloning are removed, and the library diversity is higher, and the library capacity can be as high as 1012-1014. | In vivo screening bias for certain proteins and scaffolds increases due to folding, trafficking, membrane insertion, and complexation issues, while partially unfolded proteins are often rapidly degraded within the cell. The mRNA display technology is not limited by these factors. | The screening process of mRNA display technology can be carried out under more stringent conditions, including no natural amino acid, unusual pH value and temperature, etc., and is not easily interfered by other conditions. | After the affinity screening is completed, the DNA template generated by the screening can enter the next display cycle after PCR amplification, thus greatly shortening the experimental cycle |
mRNA display can screen trillions of target molecules for desired functions in a single experiment. CD BioSciences uses this technology to build a multi-molecular discovery platform that can discover any target of interest for global customers.
The multi-application molecular discovery platform based on mRNA display technology developed by CD BioSciences will continue to provide professional services to global customers to promote drug discovery and basic research in the field of life science. At the same time, we will continue to combine multidisciplinary knowledge and continue to expand the scope of services to meet the changing research needs in the future. Please contact us to learn more about mRNA display technology and related services.
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