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Recently, biomedical engineering researchers at the University of Cincinnati have incorporated a biological pore into a lipid membrane, allowing the passage of nanomaterials from one side to the other under an applied voltage. The fundamental study was led by UC biomedical engineering Professor Peixuan Guo and the Dean of Engineering, Carlo Montemagno. Their results were published in the September 27, 2009 issue of Nature Nanotechnology ("Translocation of double-stranded DNA through membrane-adapted phi29 motor protein nanopores").
Nanopore analysis based on biological pores embedded in lipid membranes as well as solid state synthetic nanopores is currently an area of great interest in many disciplines, including cell physiology, electrical technology, and other nanotechnological applications such as single molecule sensing, molecular fingerprinting, Micro-Electro-Mechanical Systems (MEMS), microreactors, and the sequencing of DNA. The biological nanopore used in this study is a constituent of the bacteriophage phi29. One of the first challenges the team faced was “moving it from its native enclosure into this engineered environment" stated David Wendell, PhD, co-first author of the paper and Assistant Professor in UC's Biomedical and Environmental Engineering departments. To accomplish this Drs. Wendell and Montemagno developed a lipid vesicle fusion technique, based on one they previously used for ion channels studies, to embed the re-engineered nanomotor core into a lipid sheet. Once in the sheet the nanopore created a channel large enough to allow the passage of double-stranded DNA through when voltage was applied.
This work has the potential for extension in a multitude of applications, including gene delivery, drug loading and DNA sequencing. Furthermore, this system possesses a major advantage in that the dimensions of the pore are large enough to allow the translocation of dsDNA and Guo notes that "Since the genomic DNA of human, animals, plants, fungus and bacteria are double stranded, the development of single pore system that can sequence double-stranded DNA is very important.
The work in this area continues as the group studies how to apply these findings to the delivery of therapeutic agents directly to cancerous or viral-infected cells. Research continues in an effort to find the best way to use the pore to load drugs or DNA into a liposome which could then be used as a vehicle for targeted delivery. In a separate vein, Dr. Guo and his group continue to study potential sensing and sequencing applications of the system. "The idea that a DNA molecule travels through the nanopore, advancing nucleotide by nucleotide, could lead to the development of a single pore DNA sequencing apparatus, an area of strong national interest," Peng Jing, Ph.D, the other co-first author added.
Funding for this study comes from the National Institutes of Health's Nanomedicine Development Center, of which Peixuan Guo is the director.