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

Nat Commun. 2019 Oct 24;10(1):4845. doi: 10.1038/s41467-019-12800-7.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cold Temperature*
  • DNA / metabolism*
  • DNA Nucleotidyltransferases
  • Gene Regulatory Networks
  • Genetic Engineering / methods*
  • HEK293 Cells
  • Humans
  • Integrases
  • Light*
  • Mice
  • Recombinases / genetics*
  • Recombinases / metabolism

Substances

  • Recombinases
  • DNA
  • Cre recombinase
  • DNA Nucleotidyltransferases
  • FLP recombinase
  • Integrases