In Vivo, Cardiac-Specific Knockdown of a Target Protein, Malic Enzyme- 1, in Rat via Adenoviral Delivery of DNA for Non-Native miRNA
J. Michael O’Donnell,
Edward D. Lewandowski.
This study examines the feasibility of using the adenoviral delivery of DNA for a non-native microRNA to
suppress expression of a target protein (cytosolic NADP+-dependent malic-enzyme 1, ME1) in whole heart in vivo, via an
isolated-heart coronary perfusion approach. Complementary DNA constructs for ME1 microRNA were inserted into adenoviral
vectors. Viral gene transfer to neonatal rat cardiomyocytes yielded 65% suppression of ME1 protein. This viral
package was delivered to rat hearts in vivo (Adv.miR_ME1, 1013 vp/ml PBS) via coronary perfusion, using a cardiacspecific
isolation technique. ME1 mRNA was reduced by 73% at 2-6 days post-surgery in heart receiving the
Adv.miR_ME1. Importantly, ME1 protein was reduced by 66% (p<0.0002) at 5-6 days relative to sham-operated control
hearts. Non-target protein expression for GAPDH, calsequestrin, and mitochondrial malic enzyme, ME3, were all unchanged.
The non-target isoform, ME2, was unchanged at 2-5 days and reduced at day 6. This new approach demonstrates
for the first time significant and acute silencing of target RNA translation and protein content in whole heart, in vivo, via
non-native microRNA expression.
Keywords: Heart, RNA interference, gene therapy, microRNA, siRNA, heart, RNA interference, gene therapy, microRNA, siRNA, calsequestrin, mitochondrial malic enzyme, ME3
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