In this work, an intracellular protein delivery methodology termed “proteolistics” is described. This method utilizes a biolistic gun apparatus and involves a simple protein/projectile preparation step. The protein to be delivered is mixed with a gold particle microprojectile suspension and is placed onto a gene gun cartridge, where it is dehydrated using either lyophilization or room-temperature air-drying. Subsequent intracellular protein delivery is achieved in plant and mammalian tissues upon bombardment. Because the method does not require modification of delivery agents or cargo biomolecules and involves a simple physical deposition of the protein onto the microprojectiles, there is no restriction on protein type in terms of molecular weight, isoelectric point or tertiary structure. Because the method delivers protein through the widely used gene gun system, it can be readily applied to any tissue or organism amenable to biolistics. A variety of proteins with molecular weight ranging from 24 to 68 kDa and isoelectric point from 4.8 to 10.1 were tested in this work. It is anticipated that this simple and versatile technique will offer biologists a powerful tool for basic research in areas such as understanding of cell and gene functions and for biotechnological applications such as genome editing.
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S.M.-O. and K.W. thank Angela Nguyen and Xing Xu for technical support, Bronwyn Frame and Katey Warnberg for providing Hi-II maize immature ear material, Kathleen Mullin and Giuseppe Dell’Anna for providing mouse ear pinna tissue, Tracey Pepper for assistance in scanning electron microscopy imaging, and Sam Barth and Evelyn Qin for critical reading of the manuscript. This work was supported by the Plant Sciences Institute and Crop Bioengineering Consortium, Iowa State University.
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Martin-Ortigosa, S., Wang, K. Proteolistics: a biolistic method for intracellular delivery of proteins. Transgenic Res 23, 743–756 (2014). https://doi.org/10.1007/s11248-014-9807-y
- Biomolecule delivery
- Gene gun
- Protein delivery