Genome engineering with targetable nucleases (467.1)

Abstract

We are in the midst of a revolution in genome engineering. Using zinc‐finger nucleases (ZFNs), transcription activator‐like effector nucleases (TALENs), and CRISPR/Cas nucleases, researchers have made targeted genome modifications in nearly 50 different species, with remarkable efficiency. Types of targeted manipulations include local mutagenesis, specific sequence changes, large insertions and deletions, and chromosomal rearrangements. Many model organisms have been successfully targeted, as have crop plants, livestock, and disease vectors. Clinical trials in humans using these reagents are currently under way.This presentation will take a step back and look at the origins of these targetable cleavage reagents ‐ particularly the ZFNs ‐ and the DNA repair pathways they depend on for their efficacy. Specific examples of how nuclease activity has been impeded or enhanced by cell‐ and organism‐specific biology will be provided. My lab has used Drosophila to explore some of the parameters that affect nuclease efficacy. Some of our experience with ZFNs, TALENs and CRISPR/Cas will be presented, with an emphasis on issues remaining to be resolved.