Inventors at UC Irvine have engineered an orthogonal DNA replication system capable of rapid, accelerated continuous evolution. This system enables the directed evolution of specific biomolecules towards user-defined functions and is applicable to problems of protein, enzyme, and metabolic pathway engineering.
The dominant technology for directing the evolution of specific biomolecules involves rounds of in vitro PCR mutagenesis, cloning, transformation, and selection. This technique is disadvantageous because there are numerous steps and iterations, and the number of adaptive paths a biomolecule can take is limited. Further, this current approach cannot be scaled up to accommodate multiple, parallel experiments for comparative analysis. Researchers at UC Irvine have addressed these shortcomings by designing a replication system that is specific, continuous, and parallelizable. This invention enablesresearchers to evolve a gene or a collection of genes toward novel functions that require long mutational paths and large-scale replication of parallel, directed evolution experiments. This system is orthogonal, meaning that the error-prone DNA replication system does not increase the mutation rate of genomic DNA, it enables targeted mutagenesis of the genes of interest. Additionally, the method is entirely in vivo, which allows for continuous mutagenesis. Unlike other systems, this technique may be conducted in highly parallel format, owning to the small volumes required.
Inventors have developed and tested several working prototypes.