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Inducible control of gene expression with destabilized Cre

Abstract

Acute manipulation of gene and protein function in the brain is essential for understanding the mechanisms of nervous system development, plasticity and information processing. Here we describe a technique based on a destabilized Cre recombinase (DD-Cre) whose activity is controlled by the antibiotic trimethoprim (TMP). We show that DD-Cre triggers rapid TMP-dependent recombination of loxP-flanked ('floxed') alleles in mouse neurons in vivo and validate the use of this system for neurobehavioral research.

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Figure 1: TMP-dependent loxP recombination in the brain of DD-Cre mice.
Figure 2: TMP-induced synaptic silencing in the brain of DD-Cre/TeNT mice.
Figure 3: DD-Cre/TeNT mice exhibit TMP-dependent loss of recognition and spatial memory.

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Acknowledgements

We thank U. Mueller (The Scripps Research Institute (TSRI)), L. Stowers (TSRI), F. Polleux (TSRI) and D. Anderson (California Institute of Technology) for advice and discussion; T.C. Südhof (Stanford), M. Goulding (Salk Institute), U. Mueller (TSRI) and M. Shimojo (TSRI) for providing mouse strains, antibodies and expression vectors; A. Roberts, S. Kupriyanov and TSRI mouse behavioral and transgenic cores for expert technical assistance; and members of the laboratories of L. Stowers and U. Mueller for help with experiments. This study was supported in part by a US National Institutes of Health R01 grant MH085776 (A.M.), the Novartis Advanced Discovery Institute (A.M.), The Baxter Foundation (A.M.), a National Institutes of Health Predoctoral Research Service Award (R.S.) and a Helen Dorris Postdoctoral Fellowship (S.P.).

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Authors

Contributions

A.M. and R.S. conceived hypotheses and designed the experiments. R.S. generated expression constructs and characterized mutant mice. M.M. and K.B. examined TMP pharmacokinetics in the brain. K.B., S.P. and N.T.-R. contributed to imaging and behavioral analyses. T.J.W. provided DD tags and assisted with interpretation of results. A.M. wrote the manuscript.

Corresponding author

Correspondence to Anton Maximov.

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Competing interests

A.M., R.S., T.J.W. and M.M. anticipate filing a provisional patent for use of DD-Cre in genetically modified animals.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–9 (PDF 2263 kb)

Supplementary Table 1

Background and TMP-dependent recombination in the brain of DD-Cre/Ai9 mice (XLSX 50 kb)

Supplementary Table 2

ZIF268/Egr1 fluorescence intensity/neuron, Syb2-positive puncta density/mm2 and EPSP amplitude (XLSX 13 kb)

Supplementary Table 3

Locomotor activity (XLSX 75 kb)

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Sando, R., Baumgaertel, K., Pieraut, S. et al. Inducible control of gene expression with destabilized Cre. Nat Methods 10, 1085–1088 (2013). https://doi.org/10.1038/nmeth.2640

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