Electroporation of murine thoracic aortas to provide a method of localized gene delivery

Rebecca Vanbuskirk
Thoracic aortic aneurysm (TAA) disease affects about 15,000 people each year.4 However, the cellular mechanism of TAA disease is not known,5 resulting in there being a high demand for research on the condition. The Akerman and Ikonomidis laboratory has demonstrated matrix metalloproteinases (MMPs) to be large players in the disease mechanism.12 As furin has been found to modulate MMP levels, this protein subsequently plays a role in TAA progression.14 However, an effective means for localized
more » ... ansfection of vectors for these proteins into tissues has not been established.8,9 I hypothesize that we can overexpress furin in the murine thoracic aorta through electroporation. An overexpression vector for furin was generated and confirmed by transforming the vector into E. coli, purifying the double-stranded DNA (dsDNA) and running it on an ethidium bromide gel, followed by final confirmation via sequencing. Direct electroporation of the murine thoracic aorta was performed. Electrical conditions comprised of 8 square-wave pulses for 10 ms each, at 200 V. These data provide significant steps towards TAA research, as there is currently very little understanding of the molecular mechanism behind TAA development;5 by contributing to this research, we are one step closer to defining a noninvasive treatment for the disease.
doi:10.17615/6qwz-sr43 fatcat:bhunnxytwrhmfnetkjrvftzjqq