The human genome was sequenced 15 years ago. Today, thousands of sequenced genomes provide a dizzying array of new information, constituting the list of parts that define organisms, and simultaneously the programming code that dictates the functions that drive life. Tools were developed that achieve precise cleavage of single, unique DNA sequences enable scientists to edit genes by creating double-stranded breaks at any user defined sequence in the massive genomes of eukaryotes, providing means to decipher, and alter, function. A game change occurred with the discovery of CRISPR/Cas9, which effectively democratized gene editing.
To unlock the full potential of this revolution, however, our capacities to disrupt or rewrite small local elements of code by gene editing must be complemented by capabilities to efficiently dock very large DNA cargos with a wide range of functions into genomic sites, by inserting large, custom designed, multifunctional DNAs at base pair precision. Large designer DNA cargoes would carry multicomponent DNA circuitry including programmable and fine-tuneable functionalities and represent the vital interface between gene editing which is the state-of-the-art at present, and genome engineering, which is the future. This challenge remains largely unresolved to date.
The research here proposed tackles precisely this fundamental bottleneck. By bringing to bear an array of sophisticated tools, integrating unique know-how with leading synthetic, biological, chemical and biomedical assets, we will establish new technology to enable highly efficient, easy-to-use docking of custom designed very large multifunctional designer DNA cargos into mammalian genomes with base pair precision, addressing significant unmet academic, biomedical and industrial needs.
