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Principal Investigator: Sangpil Yoon
Organization: UNIVERSITY OF NOTRE DAME
Fiscal Year: 2019
Award: $248,999
Funding agency: National Institute of General Medical Sciences
Project Summary / Abstract
Dr. Sangpil Yoon is a postdoctoral scholar at the Department of Biomedical Engineering at the University of
Southern California (USC). He has developed an innovative technique for the intracellular delivery of
macromolecules using high frequency ultrasound. Dr. Yoon is a perfect candidate for NIH Pathway to
Independence Award (K99/R00) and he will smoothly transit from a mentored trainee to an independent
investigator by completing research goals and career development training plans, proposed in this award
proposal. Dr. Yoon has three mentors to provide their expertise and constructive and critical advice during his
award period. Dr. Yoon will have trainings in ultrasound and medical ultrasonic transducer development from
Dr. Kirk Shung, USC, molecular and cellular biology from Dr. Yingxiao Wang, University of California, San
Diego (UCSD), and stem cell and regenerative medicine from Dr. Qi-Long Ying, USC.
Transfection methods currently available in research laboratories and clinics are based on non-targeted and
random process such as viral-vectors and electroporation. I have developed an innovative transfection method
by focusing acoustic energy within very confined area of less than 10 μm using a very high frequency
ultrasonic transducer with a center frequency of over 150 MHz, entitled acoustic-transfection. The key
innovation is that the acoustic-transfection has the capabilities of single-cell level targeting and controlling the
size of delivered macromolecules with low cytotoxicity. The hypothesis is that the developed acoustic-
transfection can deliver various kinds of macromolecules into different cells with the peculiar capabilities that
distinguish from other transfection methods.
Understanding of signaling pathways and the activation of important molecular events during intracellular and
intercellular interactions are important because these are a basic building block to identify cell phenotypes and
some molecular events are precedent for certain disease. Based on this understanding, engineering cell fate
and cell functions using efficient gene editing with CRISPR/Cas9 and the generation of iPSCs by delivering
recombinant proteins for regenerative medicine will further advance human health. Therefore, the primary goal
of this proposal is to deliver desired molecules using acoustic-transfection into designated cells to visualize
cell-to-cell interactions and the signaling of important molecular events, to induce gene expressions, and to
demonstrate the delivery of recombinant proteins labeled with fluorescence dyes. To test hypothesis and to
achieve the goal of this proposal, three specific aims were developed. 1) I will further optimize acoustic-
transfection using macromolecules with different sizes. 2) I will deliver FRET biosensors into neighboring or
single cells to visualize molecular events under stimulation. 3) pCas9_GFP will be delivered into hESCs and
GFP expression will be observed. Recombinant proteins conjugated with Alexa 488 will be continuously
delivered into hNSCs using acoustic-transfection for 8 days to confirm short-term delivery efficacy and long-
term endogenous pluripotency. The proposed acoustic-transfection will advance transfection methods with the
safe delivery of versatile molecules into cells to better understand important molecular events in cells and
develop improved therapeutic strategies to cure diseases.
Terms: <Acoustic><Acoustics><Address><Area><Award><Biology><Biomedical Engineering><Biosensor><Blood Plasma Cell><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><California><Cas nuclease technology><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Function><Cell Interaction><Cell Process><Cell Signaling><Cell membrane><Cell physiology><Cell-to-Cell Interaction><Cells><Cellular Function><Cellular Physiology><Cellular Process><Ch'i><Characteristics><Clinic><Coloring Agents><Contrast Agent><Contrast Drugs><Contrast Media><Cytoplasmic Membrane><Data><Development><Dextrans><Disease><Disorder><Dyes><ECDGF><Echography><Echotomography><Electroporation><Endo-GF><Endothelial Cell-Derived Growth Factors><Endothelial Growth Factors><Engineering><Ensure><Event><FRET><Fluorescence><Fluorescence Resonance Energy Transfer><Frequencies><Future><Förster Resonance Energy Transfer><Gene Expression><Generations><Genes><Goals><Health><Human><Immune><Immunes><Intracellular Communication and Signaling><Investigators><Kinases><Label><Laboratory Research><Lipid Bilayers><Location><Longitudinal Studies><Lytotoxicity><Medical><Medical Ultrasound><Mentors><Methods><Modern Man><Molecular><Molecular and Cellular Biology><Monitor><NIH><National Institutes of Health><Pathway interactions><Patients><Peptides><Phenotype><Phosphotransferase Gene><Phosphotransferases><Physiologic pulse><Plasma Cells><Plasma Membrane><Plasmacytes><Play><Population><Process><Progenitor Cells><Pulse><Qi><Radiation><Radiopaque Media><Recombinant Proteins><Regenerative Medicine><Reporting><Research><Research Personnel><Research Proposals><Researchers><Resolution><Role><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Specificity><Stem cells><Subcellular Process><System><Techniques><Testing><Therapeutic><Time><Training><Transfection><Transphosphorylases><Ultrasonic Imaging><Ultrasonic Transducer><Ultrasonogram><Ultrasonography><Ultrasound Diagnosis><Ultrasound Medical Imaging><Ultrasound Test><Ultrasound transducer><United States National Institutes of Health><Universities><Viral Vector><Work><base><bio-engineered><bio-engineers><bioengineering><biological sensor><biological signal transduction><career development><cell engineering><cellular engineering><customized therapy><customized treatment><cytotoxicity><daughter cell><developmental><dextran><diagnostic ultrasound><electroporative delivery><experiment><experimental research><experimental study><gene electrotransfer><hESC><human ES cell><human ESC><human embryonic stem cell><iPS><iPSC><iPSCs><improved><individualized medicine><individualized patient treatment><individualized therapy><individualized treatment><induced pluripotent stem cell><innovate><innovation><innovative><lipid bilayer membrane><long-term study><longterm study><macromolecule><molecular imaging><molecule imaging><nano particle><nano-sized particle><nanoparticle><nanosized particle><pathway><plasmalemma><plasmid DNA><plasmocyte><pluripotency><real-time images><realtime image><screening><social role><sonogram><sonography><sound measurement><spatiotemporal><tailored medical treatment><tailored therapy><tailored treatment><treatment planning><ultrasound><ultrasound imaging><ultrasound scanning><unique treatment><voltage>