Abstract
Synthetic extracellular matrices with reversibly adjustable mechanical properties are essential for the investigation of how cells respond to dynamic mechanical cues as occurring in living organisms. One interesting approach to engineer dynamic biomaterials is the incorporation of photoreceptors from cyanobacteria or plants into polymer materials. Here, we give an overview of existing photoreceptor-based biomaterials and describe a detailed protocol for the synthesis of a phytochrome-based extracellular matrix (CyPhyGel). Using cell-compatible light in the red and far-red spectrum, the mechanical properties of this matrix can be adjusted in a fully reversible, wavelength-specific, and dose-dependent manner with high spatiotemporal control.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Watt FM, Huck WT (2013) Role of the extracellular matrix in regulating stem cell fate. Nat Rev Mol Cell Biol 14(8):467–473. https://doi.org/10.1038/nrm3620
Burdick JA, Murphy WL (2012) Moving from static to dynamic complexity in hydrogel design. Nat Commun 3:1269. https://doi.org/10.1038/ncomms2271
Rosales AM, Anseth KS (2016) The design of reversible hydrogels to capture extracellular matrix dynamics. Nat Rev Mater 1:15012. https://doi.org/10.1038/natrevmats.2015.12
Uto K, Tsui JH, DeForest CA, Kim DH (2017) Dynamically tunable cell culture platforms for tissue engineering and mechanobiology. Prog Polym Sci 65:53–82. https://doi.org/10.1016/j.progpolymsci.2016.09.004
Kloxin AM, Kasko AM, Salinas CN, Anseth KS (2009) Photodegradable hydrogels for dynamic tuning of physical and chemical properties. Science 324(5923):59–63. https://doi.org/10.1126/science.1169494
Guvendiren M, Burdick JA (2012) Stiffening hydrogels to probe short- and long-term cellular responses to dynamic mechanics. Nat Commun 3:792. https://doi.org/10.1038/ncomms1792
Zhang XL, Dong CM, Huang WY, Wang HM, Wang L, Ding D, Zhou H, Long JF, Wang TL, Yang ZM (2015) Rational design of a photo-responsive UVR8-derived protein and a self-assembling peptide-protein conjugate for responsive hydrogel formation. Nanoscale 7(40):16666–16670. https://doi.org/10.1039/c5nr05213k
Liu LM, Shadish JA, Arakawa CK, Shi K, Davis J, DeForest CA (2018) Cyclic stiffness modulation of cell-laden protein-polymer hydrogels in response to user-specified stimuli including light. Adv Biosyst 2(12):1800240. https://doi.org/10.1002/Adbi.201800240
Lyu S, Fang J, Duan T, Fu L, Liu J, Li H (2017) Optically controlled reversible protein hydrogels based on photoswitchable fluorescent protein Dronpa. Chem Commun (Camb) 53(100):13375–13378. https://doi.org/10.1039/c7cc06991j
Wu X, Huang WM, Wu WH, Xue B, Xiang DF, Li Y, Qin M, Sun F, Wang W, Zhang WB, Cao Y (2018) Reversible hydrogels with tunable mechanical properties for optically controlling cell migration. Nano Res 11(10):5556–5565. https://doi.org/10.1007/s12274-017-1890-y
Wang R, Yang Z, Luo J, Hsing IM, Sun F (2017) B12-dependent photoresponsive protein hydrogels for controlled stem cell/protein release. Proc Natl Acad Sci U S A 114(23):5912–5917. https://doi.org/10.1073/pnas.1621350114
Hörner M, Raute K, Hummel B, Madl J, Creusen G, Thomas OS, Christen EH, Hotz N, Gübeli RJ, Engesser R, Rebmann B, Lauer J, Rolauffs B, Timmer J, Schamel WWA, Pruszak J, Römer W, Zurbriggen MD, Friedrich C, Walther A, Minguet S, Sawarkar R, Weber W (2019) Phytochrome-based extracellular matrix with reversibly tunable mechanical properties. Adv Mater 31(12):e1806727. https://doi.org/10.1002/adma.201806727
Kolar K, Knobloch C, Stork H, Znidaric M, Weber W (2018) OptoBase: a web platform for molecular optogenetics. ACS Synth Biol 7(7):1825–1828. https://doi.org/10.1021/acssynbio.8b00120
Mailliet J, Psakis G, Feilke K, Sineshchekov V, Essen LO, Hughes J (2011) Spectroscopy and a high-resolution crystal structure of Tyr263 mutants of cyanobacterial phytochrome Cph1. J Mol Biol 413(1):115–127. https://doi.org/10.1016/j.jmb.2011.08.023
Hörner M, Gerhardt K, Salavei P, Hoess P, Harrer D, Kaiser J, Tabor JJ, Weber W (2019) Production of phytochromes by high-cell-density E. coli fermentation. ACS Synth Biol. https://doi.org/10.1021/acssynbio.9b00267
Berkelman TR, Lagarias JC (1986) Visualization of Bilin-linked peptides and proteins in polyacrylamide gels. Anal Biochem 156(1):194–201
Acknowledgments
This work was funded by the German Research Foundation (DFG) under the Excellence Initiative (BIOSS—EXC-294) and the Excellence Strategy (CIBSS—EXC-2189—Project ID 390939984) as well as through the European Research Council under the European Community’s Seventh Framework Programme (FP7/2007-2013)/ERC Grant agreement n° 259043-CompBioMat.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hörner, M., Hoess, P., Emig, R., Rebmann, B., Weber, W. (2020). Synthesis of a Light-Controlled Phytochrome-Based Extracellular Matrix with Reversibly Adjustable Mechanical Properties. In: Niopek, D. (eds) Photoswitching Proteins . Methods in Molecular Biology, vol 2173. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0755-8_15
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0755-8_15
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0754-1
Online ISBN: 978-1-0716-0755-8
eBook Packages: Springer Protocols