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In vivo, cardiac-specific knockdown of target protein, Malic Enzyme-1, in rat via adenoviral delivery of DNA for non-native miRNA

journal contribution
posted on 09.01.2014, 00:00 authored by J. Michael O’Donnell, Asha Kalichira, Jian Bi, E. Douglas 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 cardiac-specific 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.

Funding

NIH grants: R37 HL49244 (Lewandowski MERIT), R01 HL62702.

History

Publisher Statement

This is a copy of an article published in the Current Gene Therapy © 2012 Bentham Science Publishers. The final publication is available at http://www.benthamscience.com/cgt/index.htm

Publisher

Bentham Science Publishers

Language

en_US

issn

1566-5232

Issue date

01/12/2012

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