Developing Novel Selectable Protein Engineering Using Crispr Genome Editing

While current methods of protein and metabolic engineering can rapidly introduce many mutations into a population, their lack of selectability leads to a large amount of time and energy spent screening populations to find mutants. We aim to markedly decrease screening load by utilizing the CRISPR/Cas system in Escherichia coli to kill cells which have not acquired the desired mutations. Our goal is to combine this technique with Multiplex Automated Genome Engineering (MAGE) to create a tool for highly efficient selectable protein engineering in vivo.