Temperature-dependent Insertion of Genetic Material into Genomic DNA

Recombinant viral vectors are highly useful, especially in the production of vaccines for human pathogens. Viral vectors have undergone many revisions over the years, including attenuated versions, replication-competent versions, and versions modified to alter immune response. Each time a new vector is created, technology must be used to insert the genes of interest and select for the viruses with the recombinant vector. Current methods to ensure the survival of only the viruses having the desired recombinant vectors are cumbersome and incompatible with FDA requirements. Researchers at the Biodesign Institute of Arizona State University have developed a novel temperature dependent method to control insertion of genes using homologous recombination. The method includes a genome which contains a lethal toxic gene that is active or expressed at one temperature, leading to a lethal phenotype, but inactive or not expressed at another temperature. Viruses which contain the toxic gene could then be selected against, in favor of viruses which had replaced the toxic gene with the gene of interest. This negative selection scheme provides a superior method for isolation of recombinant vectors in a safe and highly effective manner. Potential Applications Vaccine development Cellular system analysis HTP antigen screening Studying pathogenic determinants Studying protein functions Benefits and Advantages Quick, efficient and safe selection/isolation of recombinant virus No toxic substances in the resulting recombinant virus Does not require substances that may increase the rate of mutation in the virus Does not require the use of special cell lines No materials are utilized that pose a risk to humans Dowload Original PDF For more information about the inventor(s) and their research, please see Dr. Jacob's directory webpage