Inducible Genome Modification Utilizing Engineered Ty1 Retrotransposons in Saccharomyces Cerevisiae

Tools for controllable rewiring and reengineering of organisms would be greatly useful to synthetic biology. Such tools would enable the study of synthetic gene network evolution, gene dosage and inducible genetic modification of organisms. Here we investigate engineering of retrotransposons in Saccharomyces cerevisiae. Ty1 retrotransposon is amenable for gene circuit engineering. Reengineering of these retrotransposons into a helper/donor system will allow for control over de novo production and integration of a desired genetic payload. We modify an artificial intron sequence and demonstrate successful post-transcriptional splicing using the leucine selection marker (LEU2). This intron allows for direction specific splicing and should limit gene payload expression from the donor vector. The gene payload should express only after splicing and integration into the yeast host genome. With continued induction and integration, GFP expression should increase by integer multiples, allowing for future study of copy number variation in gene networks. As the Ty1 retrotransposon is quite robust, further studies can alter the size of the gene payload and incorporate more gene networks. A selective and inducible genome modification system could assist targeted therapies and allow for easy reengineering of organisms for synthetic biology.