In this project, we will be using near infrared light-activated protein-conjugated nanoparticles to deliver functional proteins and/or siRNA to primary cells via an easily scalable, high-throughput microfluidic system available in my lab. We have already synthesized and demonstrated the protein on hollow gold nanocarrier can be successfully delivered to cells and released via pulsed NIR light directly to the cell cytoplasm (1-6). A poly-histidine tagged recombinant protein is conjugated via thiol linkers to hollow gold nanoshells. The protein-HGN are incubated with cells to internalize the carriers via endocytosis facilitated by the appropriate cell penetrating peptide. Following incubation, the HGN is activated by picosecond NIR light pulses applied during flow of the cells through a microfluidic channel. The HGN carriers adsorb the light energy, which is then converted within picoseconds into “hot electrons” that cleave the thiol bonds linking the HGN to the protein cargo. In the following nanoseconds, the HGN dissipates its heat by vaporizing a minute amount of water, forming vapor nanobubbles (7) that insulate both the cell and cargo from significant temperature changes. In this project, we will work on turning this system into a cell micro-manufacturing platform to efficiently modify the cell of choice via protein or siRNA delivery. Using this method, we can create large volumes of modified cells for use in personalized treatments of disease. Our first goal will be to use NK and T-cell modifications with the CAR protein to prime these cells for immunological treatments of cancer.
Requirements: Ph. D. in Biophysics, Chemical Engineering, Physics, Materials Science or Biochemistry.
Preferred to have previous experience in cell culture, colloid chemistry and physics, biochemistry. Position open immediately.
1. G. B. Braun, T. Friman, H. B. Pang, A. Pallaoro, T. H. de Mendoza, A. M. Willmore, V. R. Kotamraju, A. P. Mann, Z. G. She, K. N. Sugahara, N. O. Reich, T. Teesalu, E. Ruoslahti, Etchable plasmonic nanoparticle probes to image and quantify cellular internalization. Nat. Mater. 13, 904-911 (2014).
2. G. B. Braun, A. Pallaoro, G. Wu, D. Missirlis, J. A. Zasadzinski, M. Tirrell, N. O. Reich, Laser-
activated gene silencing via gold nanoshell-siRNA conjugates. ACS Nano 3, 2007–2015 (2009).
3. N. Forbes, A. Pallaoro, N. O. Reich, J. A. Zasadzinski, Rapid, Reversible Release from Thermosensitive Liposomes Triggered by Near-Infra-Red Light. Particle & Particle Systems Characterization 31, 1158- 1167 (2014).
4. X. Huang, Q. Hu, G. B. Braun, A. Pallaoro, D. P. Morales, J. A. Zasadzinski, D. O. Clegg, N. O. Reich, Light-activated RNA interference in human embryonic stem cells. Biomaterials 63, 70-79 (2015).
5. X. Huang, A. Pallaoro, G. B. Braun, D. P. Morales, M. O. Ogunyankin, J. Zasadzinski, N. O. Reich,Modular plasmonic nanocarriers for efficient and targeted delivery of cancer-therapeutic siRNA. Nano Lett 14, 2046-2051 (2014).
6. D. P. Morales, G. B. Braun, A. Pallaoro, R. W. Chen, X. Huang, J. A. Zasadzinski, N. O. Reich, Targeted Intracellular Delivery of Proteins with Spatial and Temporal Control. Molecular Pharmaceutics 12, 600-609 (2015).
7. E. Y. Lukianova-Hleb, A. N. Volkov, D. O. Lapotko, Laser Pulse Duration Is Critical For the
Generation of Plasmonic Nanobubbles. Langmuir 30, 7425-7434 (2014).