Forging Tools

Science is increasingly constrained by the limitations of available technologies. By developing improved tools and methods, we can catalyze the research of entire fields. The most useful tools expand the scope of existing or emerging technologies, enabling our accumulated expertise to address previously intractable problems.

Our tool development projects focus on novel methods of editing and regulating genomes.

Many of our projects involve the RNA-guided Cas9 nuclease, which can be directed to site-specifically bind and cut almost any sequence by simply expressing a matching RNA. This is supremely useful in nearly all organisms and essential for facile genome engineering in multicellular eukaryotes. Except for cases in which our larger goal requires it, we no longer work on direct Cas9 improvements given the intense interest from many other technology development labs. However, it is so useful that many of our other projects apply it for a variety of purposes - including building safeguards for gene drives capable of spreading genome alterations made with Cas9 through populations.

A major area of focus involves ways of multiplexing CRISPR in an evolutionarily stable manner. This is challenging because both strings of guide RNAs and natural CRISPR arrays contain highly repetitive sequences that are prone to internal recombination and rearrangement. We are exploring ways of diversifying these sequences to enable highly multiplex CRISPR.

Though now a touch dated, our Review in Molecular Systems Biology provides a comprehensive background of currently available tools and technologies for genome-scale engineering, while our Perspective in Nature Methods discusses the utility of Cas9 specifically.