High-performance chemical- and light-inducible recombinases in mammalian cells and mice
| Autor: | N. T. Hang Pham, Wilson W. Wong, Nathan Tague, Micaela Trexler, William K. J. Benman, Benjamin H. Weinberg, Maciej Walkosz, Jang Hwan Cho, Billy Law, Justin H. Letendre, Jacob Beal, Leidy D. Caraballo, Charina E. Ortega, Yash Agarwal |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Light Science Gene regulatory network General Physics and Astronomy Computational biology Biology General Biochemistry Genetics and Molecular Biology Article Genome engineering Recombinases 03 medical and health sciences chemistry.chemical_compound Mice 0302 clinical medicine Gene expression Recombinase Animals Humans Gene Regulatory Networks Mammalian gene lcsh:Science Synthetic biology 030304 developmental biology 0303 health sciences Multidisciplinary Integrases HEK 293 cells Logic gates General Chemistry DNA Small molecule Cold Temperature 030104 developmental biology HEK293 Cells chemistry Genetic circuit engineering DNA Nucleotidyltransferases lcsh:Q Genetic Engineering 030217 neurology & neurosurgery |
| Zdroj: | Nature Communications, Vol 10, Iss 1, Pp 1-10 (2019) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | Site-specific DNA recombinases are important genome engineering tools. Chemical- and light-inducible recombinases, in particular, enable spatiotemporal control of gene expression. However, inducible recombinases are scarce due to the challenge of engineering high performance systems, thus constraining the sophistication of genetic circuits and animal models that can be created. Here we present a library of >20 orthogonal inducible split recombinases that can be activated by small molecules, light and temperature in mammalian cells and mice. Furthermore, we engineer inducible split Cre systems with better performance than existing systems. Using our orthogonal inducible recombinases, we create a genetic switchboard that can independently regulate the expression of 3 different cytokines in the same cell, a tripartite inducible Flp, and a 4-input AND gate. We quantitatively characterize the inducible recombinases for benchmarking their performances, including computation of distinguishability of outputs. This library expands capabilities for multiplexed mammalian gene expression control. The availability of high performance recombinases with low basal activity and high dynamic range is limited. Here the authors present a library of over 20 orthogonal split recombinases that can be induced by small molecules, light and temperature in vivo. |
| Databáze: | OpenAIRE |
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