期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 110, 期 4, 页码 1201-1210出版社
WILEY
DOI: 10.1002/bit.24792
关键词
RNA controller; synthetic biology
资金
- National Institutes of Health [RC1GM091298]
- Defense Advanced Research Projects Agency [HR0011-11-2-0002]
- National Science Foundation
- Harvard Society of Fellows
- Department of Defense through National Defense Science and Engineering Graduate Fellowship Program
Programming genetic circuits in mammalian cells requires flexible, tunable, and user-tailored gene-control systems. However, most existing control systems are either mechanistically specific for microbial organisms or must be laboriously re-engineered to function in mammalian cells. Here, we demonstrate a ribozyme-based device platform that can be directly transported from yeast to mammalian cells in a plug-and-play manner. Ribozyme switches previously prototyped in yeast are shown to regulate gene expression in a predictable, ligand-responsive manner in human HEK 293, HeLa, and U2OS cell lines without any change to device sequence nor further optimization. The ribozyme-based devices, which exhibit activation ratios comparable to the best RNA-based regulatory devices demonstrated in mammalian cells to-date, retain their prescribed functions (ON or OFF switch), tunability of regulatory stringency, and responsiveness to different small-molecule inputs in mammalian hosts. Furthermore, we observe strong correlations of device performance between yeast and all mammalian cell lines tested (R2=0.630.97). Our unique device architecture can therefore act as a rapid prototyping platform (RPP) based on a yeast chassis, providing a well-developed and genetically tractable system that supports rapid and high-throughput screens for generating gene-controllers with a broad range of functions in mammalian cells. This platform will accelerate development of mammalian gene-controllers for diverse applications, including cell-based therapeutics and cell-fate reprogramming. Biotechnol. Bioeng. 2013; 110: 12011210. (c) 2012 Wiley Periodicals, Inc.
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