High-performance chemical- and light-inducible recombinases in mammalian cells and mice

• Published in: Nature communications Vol. 10; no. 1; pp. 4845 - 10
• Main Authors: Weinberg, Benjamin H., Cho, Jang Hwan, Agarwal, Yash, Pham, N. T. Hang, Caraballo, Leidy D., Walkosz, Maciej, Ortega, Charina, Trexler, Micaela, Tague, Nathan, Law, Billy, Benman, William K. J., Letendre, Justin, Beal, Jacob, Wong, Wilson W.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24.10.2019; Nature Publishing Group; Nature Portfolio
• Subjects: 13/109; 13/31; 42/35; 59/5; 631/553/2691; 631/553/2692; 631/553/552; 96/21; Animal models; Animals; Cold Temperature; Cytokines; Deoxyribonucleic acid; DNA; DNA - metabolism; DNA Nucleotidyltransferases; Engineering; Gene expression; Gene Regulatory Networks; Genetic Engineering - methods; Genome editing; Genomes; HEK293 Cells; High performance systems; Humanities and Social Sciences; Humans; Integrases; Light; Mammalian cells; Mammals; Mice; multidisciplinary; Organic chemistry; Recombinases - genetics; Recombinases - metabolism; Rodents; Science; Science (multidisciplinary)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-019-12800-7

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.