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Protein-based materials provide an inherently biocompatible and sustainable platform for the generation of functional materials. Translating protein properties into protein films resistant to aqueous degradation is crucial for most applications such as tissue engineering and controlled drug delivery. Current methods to stabilize protein films use three main strategies: employing the relatively limited variety of naturally self-assembling proteins, using added cross-linkers or heat curing. While the cross-linking strategy generates functionally diverse structures, unreacted additives retained in cross-linked protein films can adversely affect their final behavior. Traditional heat curing results in hydrophobic surface and loss of protein inherent properties. We demonstrate here a scalable, additive-free, fluorous media assisted thermal treatment for the fabrication of stable, hydrophilic protein films. This approach is general in terms of protein building block, retaining much of their native structure and surface properties upon heating. We demonstrate the versatility of this strategy through fabrication of antifouling coatings on complex three-dimensional surfaces. The utility of these films as biomaterials is highlighted through the generation of highly biocompatible non-fouling surfaces and regulation of cellular adhesion through choice of protein precursor. 

Abstract Macrophages are key effectors of host defense and metabolism, making them promising targets for transient genetic therapy. Gene editing through the delivery of Cas9‐ribonucleoprotein (RNP) provides multiple advantages over gene delivery–based strategies for introducing CRISPR machinery to the cell. There are, however, significant physiological, cellular, and intracellular barriers to the effective delivery of the Cas9 protein and guide RNA (sgRNA) that have to date, restricted in vivo Cas9 protein–based approaches to local/topical delivery applications. Described herein is a new nanoassembled platform featuring coengineered nanoparticles and Cas9 protein that has been developed to provide efficient Cas9‐sgRNA delivery and concomitant CRISPR editing through systemic tail‐vein injection into mice, achieving >8% gene editing efficiency in macrophages of the liver and spleen.