Axon regeneration constitutes a fundamental challenge for regenerative neurobiology, which necessitates the use of tailor-made biomaterials for controllable delivery of cells and biomolecules. An increasingly popular approach for creating these materials is to directly assemble engineered proteins into high-order structures, a process that often relies on sophisticated protein chemistry. Here, we present a simple approach for creating injectable, photoresponsive hydrogels via metal-directed assembly of His6-tagged proteins. The B12-dependent photoreceptor protein CarHC can complex with transition metal ions through an amino-terminal His6-tag, which can further undergo a sol-gel transition upon addition of AdoB12, leading to the formation of hydrogels with marked injectability and photodegradability. The inducible phase transitions further enabled facile encapsulation and release of cells and proteins. Injecting the Zn2+-coordinated gels decorated with leukemia inhibitory factor into injured mouse optic nerves led to prolonged cellular signaling and enhanced axon regeneration. This study illustrates a powerful strategy for designing injectable biomaterials.
human life. [1][2][3] To achieve such a dream, choosing natural biomaterials such as nucleic acids, [4][5][6] peptides, [7] proteins, [8,9] and polysaccharides [10] as the substitutes of fossil source-based synthetic chemicals is a promising way. Correspondingly, micro/ nanoscale fabrication on natural biomaterials receives great of importance, [11] and a spatially definable property with high precision to exactly position biomaterials on a surface is highly desirable [12][13][14][15][16][17][18] toward biodegradable and biocompatible green electronic, [19][20][21][22] optical, [23][24][25][26][27][28][29] sensor, [30] energy, [31] and environmental devices. [32] Typical examples down this road include silk fibroin, [12][13][14] polysaccharides, [10] and DNA origami-mediated lithography. [5] These approaches are part of great efforts to develop "green" lithographic processing in semiconductor industry that is eligible to lessen the exposure of workers and environment toward noxious chemical and reduce waste. [13] The adoption of these strategies allows the replacement of organic solvent with water to cast and develop a resist, accompanied with the exclusion of environmentally harmful components from material processing and fabrication. However, so far the lithographic processes based on existing natural biomaterials are difficult to be exploited in scale-up industrial application, because their limited quantities, high material, and processing cost as well as small deposition area impede the practical engineering of these biomaterials. [9] Moreover, the less-controlled variation on polymorph and polydispersity of biomaterials often influences the repeatability of results. [33] Recently, our group reported the use of phase-transited lysozyme nanofilm as the resist for photolithography and electron-beam lithography (EBL), with the cost being decreased and practical applicability being potentially demonstrated. [17] Continuous efforts along this way further require much cheaper material and simpler fabrication process for large-scale industrial uses. In this respect, we pay attention to egg white due to the following reasons. First, egg white is a very common natural biomaterial just taken from egg as our daily food, which makes it has great natural abundance with a low cost. Second, in contrast to other natural competitors usually requiring strict demands on the purity, polydispersity, and polymorph stability, egg white is a combination of multiple functional nutrients, [34] which could be directly utilized without further purification and Complex lithographic steps and the use of toxic chemicals in these processes are in conflict with a sustainable human society. Development of new inexpensive and green resist, simple alternative procedures, and nontoxic solvents is the key to move recyclable micro/nanofabrication from laboratory level to industrial application in large scale. Herein, precise control on protein fragmentation/aggregation upon photo/electron irradiation is conceived into egg white-based green resist...
Design of proteins with nonlinear topologies has emerged as a nascent branch of protein engineering, but significant applications remain to be seen. Here, we demonstrate the cellular synthesis of (SpyCatcher) 4 GFP, a 4-arm star-like protein enabled by spontaneous split GFP reconstitution, which further led to the creation of various protein networks exhibiting tunable mechanics and suitability for cell encapsulation. A derivative 4-arm star-like protein, (CarH C ) 4 GFP, resulting from the conjugation of (SpyCatcher) 4 GFP with the SpyTag-fusion CarH C photoreceptors, can undergo rapid sol-gel and gel-sol transitions in response to AdoB 12 and light, respectively. The chemo-and photo-induced phase transitions enabled encapsulation and controlled release of protein molecules such as the biofilm-degrading glycosyl hydrolase PslG, a potential agent for combatting multidrug-resistant bacterial species in chronic infections. The creation of those uncommon protein architectures promises great opportunities for materials biology and ''smart'' therapeutic delivery.
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