Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Mammalian synthetic circuits with RNA binding proteins for RNA-only delivery


Synthetic regulatory circuits encoded in RNA rather than DNA could provide a means to control cell behavior while avoiding potentially harmful genomic integration in therapeutic applications. We create post-transcriptional circuits using RNA-binding proteins, which can be wired in a plug-and-play fashion to create networks of higher complexity. We show that the circuits function in mammalian cells when encoded in modified mRNA or self-replicating RNA.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: RNA-only multi-input microRNA classifier circuit differentiates between HeLa, HEK293 and MCF7 cells.
Figure 2: Post-transcriptional cascades and two-state switch.

Similar content being viewed by others


  1. Tavernier, G. et al. J. Control. Release 150, 238–247 (2011).

    Article  CAS  Google Scholar 

  2. Sahin, U., Karikó, K. & Türeci, Ö. Nat. Rev. Drug Discov. 13, 759–780 (2014).

    Article  CAS  Google Scholar 

  3. Khalil, A.S. & Collins, J.J. Nat. Rev. Genet. 11, 367–379 (2010).

    Article  CAS  Google Scholar 

  4. Aubel, D. & Fussenegger, M. BioEssays 32, 332–345 (2010).

    Article  CAS  Google Scholar 

  5. An, C.I. RNA 12, 710–716 (2006).

    Article  CAS  Google Scholar 

  6. Culler, S.J., Hoff, K.G. & Smolke, C.D. Science 330, 1251–1255 (2010).

    Article  CAS  Google Scholar 

  7. Ausländer, S. et al. Nat. Methods 11, 1154–1160 (2014).

    Article  Google Scholar 

  8. Rodrigo, G., Landrain, T.E. & Jaramillo, A. Proc. Natl. Acad. Sci. USA 109, 15271–15276 (2012).

    Article  CAS  Google Scholar 

  9. Green, A.A., Silver, P.A., Collins, J.J. & Yin, P. Cell 159, 925–939 (2014).

    Article  CAS  Google Scholar 

  10. Qian, L. & Winfree, E. Science 332, 1196–1201 (2011).

    Article  CAS  Google Scholar 

  11. Saito, H. et al. Nat. Chem. Biol. 6, 71–78 (2010).

    Article  CAS  Google Scholar 

  12. Ausländer, S., Ausländer, D., Müller, M., Wieland, M. & Fussenegger, M. Nature 487, 123–127 (2012).

    Article  Google Scholar 

  13. Van Etten, J. et al. J. Biol. Chem. 287, 36370–36383 (2012).

    Article  CAS  Google Scholar 

  14. Xie, Z., Wroblewska, L., Prochazka, L., Weiss, R. & Benenson, Y. Science 333, 1307–1311 (2011).

    Article  CAS  Google Scholar 

  15. Strauss, J.H. & Strauss, E.G. Microbiol. Rev. 58, 491–562 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Gardner, T.S., Cantor, C.R. & Collins, J.J. Nature 403, 339–342 (2000).

    Article  CAS  Google Scholar 

  17. Kramer, B.P. et al. Nat. Biotechnol. 22, 867–870 (2004).

    Article  CAS  Google Scholar 

  18. Mortimer, I. et al. Gene Ther. 6, 403–411 (1999).

    Article  CAS  Google Scholar 

  19. Azizgolshani, O., Garmann, R.F., Cadena-Nava, R., Knobler, C.M. & Gelbart, W.M. Virology 441, 12–17 (2013).

    Article  CAS  Google Scholar 

  20. Lustig, S. et al. J. Virol. 62, 2329–2336 (1988).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Frolov, I. et al. J. Virol. 73, 3854–3865 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Beal, J. et al. ACS Synth. Biol. 4, 48–56 (2015).

    Article  CAS  Google Scholar 

  23. Petrakova, O. et al. J. Virol. 79, 7597–7608 (2005).

    Article  CAS  Google Scholar 

  24. Szymczak, A.L. et al. Nat. Biotechnol. 22, 589–594 (2004).

    Article  CAS  Google Scholar 

  25. Stewart, S.A. et al. RNA 9, 493–501 (2003).

    Article  CAS  Google Scholar 

  26. MATLAB and Statistics Toolbox Release 2013b, The MathWorks, Inc., Natick, Massachusetts, USA.

Download references


We thank K. Hayashi and N. Nishimura (Kyoto University) for supporting modified mRNA experiments. We also thank A.C. Goldstrohm and C. Weidmann (University of Michigan Medical School) for sharing Drosophila constructs containing MS2-fused repressors and the corresponding reporter, X. Zhang (MIT) and O. Andries (Ghent University) for assisting in VEE replicon construction, T.E. Wagner, D. Densmore (Boston University) and S. Payne (MIT) for communicating results before publication, W.M. Gelbart, C.M. Knobler, A. Berk, O. Azizgolshani and J.M. Parker (University of California, Los Angeles) for sharing replicon constructs and expertise, as well as A. Graziano (MIT) for supporting pDNA experiments and H. Chung (Harvard University) for designing the intronic mKate construct. This work was supported by Defense Advanced Research Projects Agency DARPA-BAA-11-23, US National Institutes of Health grants no. P50-GM098792, 5-R01-CA155320-03, National Science Foundation GRFP grant no. 1122374 (R.W.) and JSPS KAKENHI grant numbers 23681042, 24104002, and Research Center Network for Realization of Regenerative Medicine from the Japan Science and Technology Agency (H.S.).

Author information

Authors and Affiliations



L.W. and R.W. conceived the ideas, L.W. designed and performed pDNA experiments, K.E. designed and performed modRNA experiments, T.K. designed and performed replicon experiments, V.S. designed and performed pDNA apoptotic assay and 3′ UTR repressor test in HeLa cells. B.S. created computational model of pDNA and replicon-based switch. L.W., R.W., K.E. and H.S. wrote the manuscript with input from all other authors.

Corresponding authors

Correspondence to Hirohide Saito or Ron Weiss.

Ethics declarations

Competing interests

H.S., K.E., R.W., L.W., T.K. and V.S. are co-inventors on patent applications covering the RNA circuits described here.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–28, Supplementary Tables 1–6 and Supplementary Notes 1–3 (PDF 14585 kb)

Supplementary Source Code

RNA circuits model (TXT 22 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wroblewska, L., Kitada, T., Endo, K. et al. Mammalian synthetic circuits with RNA binding proteins for RNA-only delivery. Nat Biotechnol 33, 839–841 (2015).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research