High-performance chemical- and light-inducible recombinases in mammalian cells and mice.
High-performance chemical- and light-inducible recombinases in mammalian cells and mice.
复制标题
哺乳动物细胞和小鼠中的高性能化学和光诱导重组酶。
DOI：
10.1038/s41467-019-12800-7
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发表时间：
2019
影响因子：
16.6
通讯作者：
Wong,WilsonW
中科院分区：
文献类型：
--
作者：
Weinberg,BenjaminH;Cho,JangHwan;Agarwal,Yash;Pham,NTHang;Caraballo,LeidyD;Walkosz,Maciej;Ortega,Charina;Trexler,Micaela;Tague,Nathan;Law,Billy;Benman,WilliamKJ;Letendre,Justin;Beal,Jacob;Wong,WilsonW
位点特异性DNA重组酶是重要的基因组工程工具。特别是化学和光诱导的重组酶，能够时空控制基因表达。然而，由于工程化高性能系统的挑战，诱导型重组酶是稀缺的，从而限制了可以创建的遗传电路和动物模型的复杂性。在这里，我们提出了一个包含>20种正交诱导型分裂重组酶的文库，这些重组酶可以在哺乳动物细胞和小鼠中被小分子、光和温度激活。此外，我们的工程诱导分裂Cre系统具有更好的性能比现有的系统。使用我们的正交诱导重组酶，我们创建了一个遗传开关板，可以独立地调节3种不同的细胞因子在同一细胞中的表达，一个三方诱导Flp，和一个4输入与门。我们定量地表征了诱导型重组酶的基准性能，包括计算输出的可重复性。该文库扩展了多重哺乳动物基因表达控制的能力。
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.