mRNA delivery is a promising avenue for the application of novel gene therapies. The expression of therapeutic proteins from exogenous DNA requires transport of these constructs to the nucleus for transcription, complicating the route to expression. Additionally, current methods of DNA delivery, such as lentiviral vectors, come with the risk of random genome integration which can induce dangerous mutations in transduced cells.

Delivering therapeutic proteins encoded in mRNA is not without drawbacks. Cell cultures treated with untreated mRNA exhibit an inflammatory response when compared to cells treated with encapsulated mRNA (Uchida et al., 2013). We think V.I.N.C.En.T. could help with this!

In order to show the capacity for delivery of mRNA of one of our most interesting PNCs, we attempted to deliver an mRNA encoding a green fluorescent protein called Clover to cultures of HT22 neuronal cells using our minimal Arc Gag particles. This protein was designed by selecting the region of the Arc coding sequence with the highest homology to the HIV-1 Gag protein. It has been theorized that the stem loops in the RNA sequence encoding the N-lobe of the HIV-1 Gag protein play a role in regulating the uptake of RNA into the Gag complex, so we also investigated the effect of adding this sequence to the 3’ untranslated region of the Clover mRNA.

The spread of invasive Dreissena mussel species, especially Zebra (D. polymorpha) and Quagga (D. bugensis) mussels, across aquatic environments in Canada represents serious ecological and economic hazard. Originating in Russia and the Ukraine, these mussel species spread rapidly through lakes and rivers to which they are introduced. They are responsible for injuries to swimmers, mass die-offs of native mollusk species, and hundreds of millions of dollars per year in damage to industries dependent on the Great Lakes (Rosaen et al. 2012). While Dreissena species have not yet invaded Alberta, the impact of a significant infestation would be felt across a wide range of industries.

One of the most promising new methods of controlling the spread of invasive mussels is Zequanox®, a cell lysate of Pseudomonas fluorescens. This treatment is believed to be effective due to FitD, a protein found in P. fluorescens that exhibits molluscicidal activity specifically against Dreissena species while sparing most indigenous mollusk species.

To demonstrate that PNCs enable a wide range of applications, including species-specific control of invasive organisms that pose an economic or environmental threat, we designed a PNC based system for delivering the protein toxin, FitD, to mussels. We chose to base our design on the bacteriophage P22 coat protein, as this PNC has well defined methods for encapsulating protein cargo and decorating the surface with functional peptides. See our Product Design page for more information

In collaboration with the University of Calgary....

• Department of Fisheries and Oceans Canada. (2014). Lake Winnipeg Zebra Mussel treatment. DFO Canada Science Advisory Secretariat Science Response. 2014/031
• Uchida, S., Itaka, K., Uchida, H., Hayakawa, K., Ogata, T., Ishii, T., Fukushima, S., Osada, K., Kataoka, K. (2013) In Vivo Messenger RNA Introduction into the Central Nervous System Using Polyplex Nanomicelle. PLOS ONE, 8, e56220.