Biology and Application of Genome Editing

Precision genome editing, which can be used to alter specific DNA sequences, is a powerful tool for understanding the molecular circuitry underlying cellular processes. Over the past several years, we and others have harnessed microbial CRISPR-Cas systems for use as platforms for a range of genome manipulations, including single and multiplex gene knockout, gene activation, and large-scale screening applications. Recently, we discovered and characterized several novel CRISPR systems that target RNA, including the CRISPR-Cas13 family. We developed a toolbox for RNA modulation based on Cas13, including methods for highly specific RNA knockdown, transcript imaging, and precision base editing. During our initial characterization of Cas13, we observed that Cas13 also exhibits so-called non-specific "collateral" RNase activity in vitro, which we capitalized on to create SHERLOCK, a highly sensitive and specific CRISPR diagnostic platform. We are continuing to refine and extend CRISPR-based technologies as well as explore microbial diversity to find new enzymes and systems that can be adapted for use as molecular biology tools and novel therapeutics.