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NameMr. Noah Alter
Organization or InstitutionNova Southeastern University’s Kiran C. Patel College of Allopathic Medicine
TopicBiochemistry / Chem Bio.
Title

Assembly of nanoparticle-peptide vehicles for stem cell gene transfection.

Author(s)

Noah Alter, Katelyn Carnevale

Author Institution(s)

Nova Southeastern University’s Kiran C. Patel College of Allopathic Medicine

Abstract

Solid-state nanoparticles (gold, silica, or semiconductor CdSe@ZnS), biofunctionalized with nucleic acids (DNA or RNA) represent a highly promising avenue in the field of targeted biomedical therapies due to their tunable physiochemical properties and the possibility of producing modulable therapies specific to disease states. Combing this biochemical tool with cell-penetrating peptides (CPPs), which are peptide sequences known to promote cellular entry, creates a versatile cell transfection and therapy platform. Primary cell populations, such as human mesenchymal stem cells (hMSCs) isolated from bone marrow, can be difficult to transfect. Identifying which CPP would be the most advantageous transfection agent in hMSCs is useful for the development of efficacious DNA-peptide-nanoparticle constructs. These constructs can be used to generate functional hMSC-based bioreactors, capable of producing a variety of therapeutic proteins, which could then be reintroduced to treat a patient, after efficient cellular modification. Once assembled with a screen of CPP sequences to assist with hMSC cellular entry and DNA plasmids to promote fluorescent gene expression within the stem cells, the DNA-peptide-nanoparticle constructs’ induced gene expression can be observed and compared by live-cell fluorescence microscopy. Fluorescence imaging is used to quantify the fluorescence intensity and timing of protein expression to determine the optimum CPP for hMSC transfection and efficient gene expression of the fluorescent protein gene, as a model of therapeutic DNA. This approach allows for the potential genetic engineering of cellular bioreactors capable of producing tailored gene therapy for any disease with known genetic etiology.