TY - JOUR
T1 - The COMET toolkit for composing customizable genetic programs in mammalian cells
AU - Donahue, Patrick S.
AU - Draut, Joseph W.
AU - Muldoon, Joseph J.
AU - Edelstein, Hailey I.
AU - Bagheri, Neda
AU - Leonard, Joshua N.
N1 - Funding Information:
This work was supported in part by the National Cancer Institute (NCI) of the National Institutes of Health (NIH) under Award Number F30CA203325; the National Institute of Biomedical Imaging and Bioengineering of the NIH under Award Number 1R01EB026510; the National Institute of General Medical Sciences (NIGMS) of the NIH under Award Number T32GM008152 (to Hossein Ardehali); the Northwestern University Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI 5P30CA060553); the NUSeq Core of Northwestern’s Center for Genetic Medicine, and a seed grant from the Northwestern Institute for Cellular Engineering Technologies (iCET). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The authors would like to thank Everett Allchin, Lauren Battaglia, Pear Dhiantravan, Benjamin Leibowitz, Sophia Li, Siyuan Feng, and Viswajit Kandula for their assistance with cloning in this paper, and Amy Hong, Cameron McDonald, Justin Finkle, and Sebastian Bernasek for useful discussion on the computational model. The authors thank Ahmad Khalil (Boston University) for sharing plasmids encoding the ZFa from his 2012 study15.
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Engineering mammalian cells to carry out sophisticated and customizable genetic programs requires a toolkit of multiple orthogonal and well-characterized transcription factors (TFs). To address this need, we develop the COmposable Mammalian Elements of Transcription (COMET)—an ensemble of TFs and promoters that enable the design and tuning of gene expression to an extent not, to the best of our knowledge, previously possible. COMET currently comprises 44 activating and 12 inhibitory zinc-finger TFs and 83 cognate promoters, combined in a framework that readily accommodates new parts. This system can tune gene expression over three orders of magnitude, provides chemically inducible control of TF activity, and enables single-layer Boolean logic. We also develop a mathematical model that provides mechanistic insights into COMET performance characteristics. Altogether, COMET enables the design and construction of customizable genetic programs in mammalian cells.
AB - Engineering mammalian cells to carry out sophisticated and customizable genetic programs requires a toolkit of multiple orthogonal and well-characterized transcription factors (TFs). To address this need, we develop the COmposable Mammalian Elements of Transcription (COMET)—an ensemble of TFs and promoters that enable the design and tuning of gene expression to an extent not, to the best of our knowledge, previously possible. COMET currently comprises 44 activating and 12 inhibitory zinc-finger TFs and 83 cognate promoters, combined in a framework that readily accommodates new parts. This system can tune gene expression over three orders of magnitude, provides chemically inducible control of TF activity, and enables single-layer Boolean logic. We also develop a mathematical model that provides mechanistic insights into COMET performance characteristics. Altogether, COMET enables the design and construction of customizable genetic programs in mammalian cells.
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U2 - 10.1038/s41467-019-14147-5
DO - 10.1038/s41467-019-14147-5
M3 - Article
C2 - 32034124
AN - SCOPUS:85079083753
VL - 11
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 779
ER -