The Telomerator: a new tool for chromosome engineering

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Leslie Mitchell, Jef D. Boeke

Johns Hopkins University School of Medicine, United States

Chromosome engineering is an emerging focus in the fields of systems biology, genetics, synthetic biology, and the functional analysis of genomes. Here we describe the ‘telomerator’, a new technology/synthetic biology “device” designed to inducibly linearize circular DNA molecules in vivo in Saccharomyces cerevisiae. From a basic science perspective, this tool offers a new way to study the effect of gene placement on chromosomes (i.e. telomere proximity), the essentiality of 3’ non-coding regions of genes, and the plasticity of gene order and chromosome structure on cell fitness. Commercially, this tool provides a flexible new strategy to aid in the construction and expression of large, non-native pathways in S. cerevisiae on supernumerary chromosomes. We demonstrate the function of the telomerator by circularly permuting synIXR, a synthetic yeast chromosome arm previously shown to power growth of a yeast cell as a circular DNA molecule. The resulting 53 permuted strains exhibit differential growth patterns, indicating telomerator-driven linearization is a new way to generate phenotypic diversity. In principle this technology can be extended to other eukaryotic organisms.