in vivo gene silencing with siRNA using Magnetofection with in vivo PolyMag

30 µg of siRNA were mixed with 30 µL of in vivo PolyMag to efficiently target gene silencing into left adductor muscle of mice.

This article demonstres the capacity of Magnetofection technology to efficiently silence gene expression in vivo with siRNA using in vivo PolyMag from OZ Biosciences.

article reference: J Biol Chem. 2014 Apr 6.

Neuron-derived neurotrophic factor functions as a novel modulator that enhances endothelial cell function and revascularization processes.

Ohashi K, Enomoto T, Joki Y, Shibata R, Ogura Y, Kataoka Y, Shimizu Y, Kambara T, Uemura Y, Yuasa D, Matsuo K, Hayakawa S, Hiramatsu-Ito M, Murohara T, Ouchi N.

Abstract

Strategies to stimulate revascularization are valuable for cardiovascular diseases. Here we identify neuron-derived neurotrophic factor (NDNF)/epidermacan as a secreted molecule that is upregulated in endothelial cells in ischemic limb of mice. NDNF was secreted from cultured human endothelial cells, and its secretion was stimulated by hypoxia. NDNF promoted endothelial cell network formation and survival in vitro through activation of Akt/eNOS signaling involving integrin αvβ3. Conversely, siRNA-mediated knockdown of NDNF in endothelial cells led to reduction of cellular responses and basal Akt signaling. Intramuscular overexpression of NDNF led to enhanced blood flow recovery and capillary density in ischemic limb of mice, which was accompanied by enhanced phosphorylation of Akt and eNOS. The stimulatory actions of NDNF on perfusion recovery in ischemic muscle of mice were abolished by eNOS deficiency or NOS inhibition. Furthermore, siRNA-mediated reduction of NDNF in muscle of mice resulted in reduction of perfusion recovery and phosphorylation of Akt and eNOS in response to ischemia. Our data indicate that NDNF acts as an endogenous modulator that promotes endothelial cell function and ischemia-induced revascularization through eNOS-dependent mechanisms. Thus, NDNF can represent a therapeutic target for the manipulation of ischemic vascular disorders.

In vivo PolyMag from OZ Biosciences is a very efficient delivery reagent based on Magnetofection™ technology for targeting gene silencing in vivo.