@article {Huang127068, author = {Yuping Huang and Caitlin McCann and Andrey Samsonov and Dmitry Malkov and Gregory D Davis and Qingzhou Ji}, title = {Modulation of Genome Editing Outcomes by Cell Cycle Control of Cas9 Expression}, elocation-id = {127068}, year = {2017}, doi = {10.1101/127068}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Targeting specific chromosomal sequences for genome modification or regulation during particular phases of the cell cycle may prove useful in creating more precise, predictable genetic changes. Here, we present a system using a fusion protein comprised of a programmable DNA modification protein, Cas9, linked to a cell cycle regulated protein, geminin, as well as green fluorescent protein (GFP) for visualization. Despite the large size of Cas9 relative to geminin, cells were observed to express Cas9-GFP-geminin at levels which oscillate with the cell cycle. These fusion proteins are also shown to retain double-strand break (DSB) activity at specific chromosomal sequences to produce both indels and targeted integration of donor ssDNA. Most importantly, the ratio of ssDNA donor integration to non-homologous end joining (NHEJ) was observed to increase, suggesting that cell cycle control Cas9 expression may be an effective strategy to bias DNA repair outcomes.}, URL = {https://www.biorxiv.org/content/early/2017/04/12/127068}, eprint = {https://www.biorxiv.org/content/early/2017/04/12/127068.full.pdf}, journal = {bioRxiv} }