CRISPR-Mediated Genetic Diversification in Yeast
Directed evolution takes advantage of repeated Darwinian cycles of genetic diversification and artificial selection to engineer novel biomolecular and cellular function. In the wet-lab, its success and extent is heavily dictated by the size of the genetic library that can be synthesized and transformed, in combination with labor, cost, and time. Therefore, traditional methods relying on ex-vivo diversification and in-vivo selection are highly limiting. By addressing these problems, methods for targeted, continuous in-vivo mutagenesis are extremely valuable. Recently, our lab group developed one such system termed EvolvR – an HDR-independent, multiplexable, targeted genetic diversifier that localizes error prone DNA polymerases to user-defined loci via CRISPR-guided Cas9-nickases. Prior work was carried out in E. coli; however, EvolvR holds immense promise as a species-independent technology whereby genes of interest can be evolved in a native or more suitable context. Thus, translating EvolvRs utility to yeast holds vast potential in directing the evolution of new biotechnologically useful phenotypes and paves the way for its further translation to other eukaryotic cells types.
Message to Sponsor
- Major: Bioengineering
- Sponsor: Rose Hills Independent
- Mentor: John Dueber