Possible applications for this invention include transient expression of specific gene products in stem cells prior to transplantation to protect the cells or stimulate them to differentiate into a desired cell-type (e.g., pancreatic islet cells for diabetic patients). This technology could also have great utility in the development of RNA-based vaccines and gene-based therapies for different forms of cancer.
In eukaryotes, protein synthesis is the process by which messenger RNA (mRNA) interacts with ribosomes, cofactors, and enzymes to translate the mRNA's nucleotide sequence to an amino acid sequence. The translation process is complex and can be modified at a number of steps. For example, the initiation of translation requires a critical interaction between the mRNA and specific binding proteins called initiation factors. An important initiation factor in mammalian cells is eukaryotic Initiation Factor-4E (eIF-4E). Translation is further regulated by other factors and conditions, including protein that specifically bind initiation factors. Recent studies have shown that eIF-4E appears to be directly involved in the maintenance and expression of certain forms of cancer. Dysregulation eIF-4E may also be involved in other disease states as well (e.g., diabetes).
This technology provides recombinant human eIF-4E and a method for purifying it from expressing cells. It also provides methods for the expression of eIF-4E with amino acid sequence variants, which alter its RNA binding affinity. These methods can be used both in vivo and in vitro for the investigation of cancer and other disease states.