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New Research Holds Promise for Gene Engineering and Therapy
Date:4/4/2011

terial a scientist needed for creating proteins from a cell's nuclear machinery. But virus bodies don't work terribly well because cells recognize them for what they are...or were: viruses. When host cells sense the presence of these viral guests they sound immunological alarms.

The most popular alternatives to viral vehicles are lipids and cell permeating peptides (CPP), which do a similar job without inciting a cell to rebellion. Yamano's and Montclare's achievement was to create a hybrid comprising two of these: the well-known transfection reagent FuGENE HD (FH), which is a lipid construct, and a modified version of the oft-used CPP HIV-1 Tat (mTat).

Yamano, a former fellow at the National Institutes of Health and Harvard whose research focuses on gene therapeutics for oral diseases, says he arrived at the idea to combine the two transfection vehicles because he knew FH by itself is the best stand-alone transfection reagent across a range of cell lines. He also reasoned that mTat, when modified with histidine and cysteine residues, works better than the unmodified CPP Tat. Finally, and perhaps most critically, while mTat does not transfect in a serum medium, which is basically the natural milieu of animal and human cells, FH does – and particularly well.

Thus the researchers set out to create the best of both worlds by combining FH and mTat. After all, if FH can work in a serum medium, perhaps mTat "wearing" FH as a kind of molecular raincoat could, too. With her prior research into protein engineering, Montclare would be a natural to perform the physical characterization of these complexes. Her work helped Yamano's lab understand surface charges and other subtleties of the new complex.

Indeed, Yamano and Montclare found that on five different cell types, the new mTat/FH complex was about four times as effective in a medium with serum than FH alone. "This result suggests our vector may have a great potential clinical application
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SOURCE Polytechnic Institute of New York University
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