Abstract
The ability to apply precise inputs to signaling species in live cells would be transformative for interrogating and understanding complex cell-signaling systems. Here we report an 'optogenetic' method for applying custom signaling inputs using feedback control of a light-gated protein-protein interaction. We applied this strategy to perturb protein localization and phosphoinositide 3-kinase activity, generating time-varying signals and clamping signals to buffer against cell-to-cell variability or changes in pathway activity.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Mettetal, J.T., Muzzey, D., Gomez-Uribe, C. & van Oudenaarden, A. Science 319, 482–484 (2008).
Muzzey, D., Gómez-Uribe, C.A., Mettetal, J.T. & van Oudenaarden, A. Cell 138, 160–171 (2009).
Shimizu, T.S., Tu, Y. & Berg, H.C. Mol. Syst. Biol. 6, 382 (2010).
Hodgkin, A.L., Huxley, A.F. & Katz, B. J. Physiol. (Lond.) 116, 424–448 (1952).
Visscher, K., Schnitzer, M.J. & Block, S.M. Nature 400, 184–189 (1999).
Rockwell, N.C., Su, Y.S. & Lagarias, J.C. Annu. Rev. Plant Biol. 57, 837–858 (2006).
Khanna, R. et al. Plant Cell 16, 3033–3044 (2004).
Levskaya, A., Weiner, O.D., Lim, W.A. & Voigt, C.A. Nature 461, 997–1001 (2009).
Shimizu-Sato, S., Huq, E., Tepperman, J.M. & Quail, P.H. Nat. Biotechnol. 20, 1041–1044 (2002).
Tyszkiewicz, A.B. & Muir, T.W. Nat. Methods 5, 303–305 (2008).
Leung, D.W., Otomo, C., Chory, J. & Rosen, M.K. Proc. Natl. Acad. Sci. USA 105, 12797–12802 (2008).
Edelstein, A., Amodaj, N., Hoover, K., Vale, R. & Stuurman, N. Curr. Protoc. Mol. Biol. 92, 14.20.01–14.20.17 (2010).
Suh, B.C., Inoue, T., Meyer, T. & Hille, B. Science 314, 1454–1457 (2006).
Haugh, J.M., Codazzi, F., Teruel, M. & Meyer, T. J. Cell Biol. 151, 1269–1280 (2000).
Gibson, D.G. et al. Nat. Methods 6, 343–345 (2009).
Ory, D.S., Neugeboren, B.A. & Mulligan, R.C. Proc. Natl. Acad. Sci. USA 93, 11400–11406 (1996).
Toettcher, J.E., Gong, D., Lim, W.A. & Weiner, O.D. Methods Enzymol. 497, 409–423 (2011).
Acknowledgements
We thank all members of the Weiner and Lim laboratories, A. Levskaya, A. Edelstein and N. Stuurman for helpful comments and discussions, and A. Loewer and H. El-Samad for critically reading this manuscript. TagBFP construct was a gift from members of the Vale lab (University of California San Francisco), pHR lentiviral vectors were provided by J.R. James (University of California San Francisco), and the iSH construct was a gift from members of the Meyer lab (Stanford University). This work was supported by a Cancer Research Institute Postdoctoral Fellowship to J.E.T., an American Cancer Society postdoctoral fellowship to D.G., the Howard Hughes Medical Institute and US National Institutes of Health grants GM55040, GM62583, EY016546 and P50GM081879 to W.A.L., and a National Institutes of Health grant GM084040 and a Searle Scholars Fellowship to O.D.W.
Author information
Authors and Affiliations
Contributions
J.E.T. conceived and implemented the feedback controller. O.D.W. and W.A.L. supervised the project. All authors designed experiments, which J.E.T. and D.G. performed. J.E.T. analyzed data. J.E.T., W.A.L. and O.D.W. wrote the paper, which all authors edited.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–9, Supplementary Note 1 (PDF 1401 kb)
Supplementary Software 1
Matlab toolbox for microscope control (via Micro-Manager) and the feedback controller. (ZIP 28 kb)
Rights and permissions
About this article
Cite this article
Toettcher, J., Gong, D., Lim, W. et al. Light-based feedback for controlling intracellular signaling dynamics. Nat Methods 8, 837–839 (2011). https://doi.org/10.1038/nmeth.1700
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nmeth.1700
This article is cited by
-
Maximizing protein production by keeping cells at optimal secretory stress levels using real-time control approaches
Nature Communications (2023)
-
High-throughput feedback-enabled optogenetic stimulation and spectroscopy in microwell plates
Communications Biology (2023)
-
Optimal control of bioproduction in the presence of population heterogeneity
Journal of Mathematical Biology (2023)
-
Microscopes are coming for your job
Nature Methods (2022)
-
CyberSco.Py an open-source software for event-based, conditional microscopy
Scientific Reports (2022)