A GFP complementation system for monitoring and directing nanomaterial mediated protein delivery to human cellular organelles

Shyam Sundhar Bale, Seok Joon Kwon, Dhiral A. Shah, Ravi S. Kane, Jonathan S. Dordick

Research output: Contribution to journalLetterpeer-review

13 Scopus citations


Protein-based therapeutics are gaining importance for their biocompatibility and activity toward specific targets. When these targets are intracellular, it is critical to deliver biomolecules to sites in the cell cytoplasm while retaining biomolecule activity in the complex cellular milieu. However, intracellular protein delivery is not viable unless accompanied by an active uptake mechanism or carrier mediated delivery. Moreover, once entry into the cell is achieved, detection of the biomolecule requires laborious techniques that lack real-time measurement. We have developed a fluorescence-based complementary protein delivery sensing system using split green fluorescence protein (GFP1-10 and GFP11) fragments, which can be used as an indicator for protein delivery and retention of activity, and as a means to pinpoint subcellular localization. We demonstrate in vitro localized delivery by expressing the GFP11 fragment onto the mitochondrial outer membrane of human cells, and using a model carrier (15 nm silica nanoparticles) to deliver GFP1-10 and image trafficking and mitochondrial localization of protein delivery. Our results indicate that nanoscale materials can be used as protein carriers for targeting cell constituents including functional molecules, signaling pathways, and organelles. We envision that this GFP complementation system is ideally suited for directing nanoparticle-based delivery of drugs and other bioactive molecules into subcellular locations within cells, which can impact protein-protein interactions, signal transduction pathways, and organelle function in vitro within the context of high-throughput screening protocols.

Original languageEnglish
Pages (from-to)1040-1047
Number of pages8
JournalBiotechnology and Bioengineering
Issue number6
StatePublished - 15 Dec 2010
Externally publishedYes


  • GFP complementation
  • Mitochondrial delivery
  • Nanoparticle-protein delivery
  • Protein delivery sensor
  • Sub-cellular localization


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