alexa miRNA Lentiviral Vector Integration and Gene Targeting Efficacy in Cardiac Progenitors
ISSN: 2157-7633

Journal of Stem Cell Research & Therapy
Open Access

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Research Article

miRNA Lentiviral Vector Integration and Gene Targeting Efficacy in Cardiac Progenitors

Mariana Loperfido1, Stefania Crippa1 and Maurilio Sampaolesi1,2*

1Translational Cardiomyology, Stem Cell Research Institute, KULeuven, Herestraat 49, 3000 Leuven, Belgium

2Human Anatomy, University of Pavia, Via Forlanini 8, 27100 Pavia, Italy

*Corresponding Author:
Maurilio Sampaolesi, Ph.D
Stem Cell Research Institute, KULeuven
Herestraat 49, 3000 Leuven, Belgium
Tel: +32-(0)163-30295
Fax: +32-(0)163-30294
E-mail: [email protected]

Received date May 08, 2012; Accepted date June 27, 2012; Published date June 29, 2012

Citation: Loperfido M, Crippa S, Sampaolesi M (2012) miRNA Lentiviral Vector Integration and Gene Targeting Efficacy in Cardiac Progenitors. J Stem Cell Res Ther S9:003. doi:10.4172/2157-7633.S9-003

Copyright: © 2012 Loperfido M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Stem and progenitor cardiac cells are challenging for possible cell therapy application. Several research laboratories are exploiting the feasibility of autologous cell therapy approach to get rid of immunosuppressive treatments responsible for undesirable side effects. Recently, we showed that cardiac progenitors isolated from Sgcb null mice, animal model of limb-girdle muscular dystrophy type 2E, undergo an aberrant differentiation in vitro and in vivo due to the dysregulation of miR669. This miRNA family is able to inhibit the skeletal myogenic program directly targeting MyoD 3’ UTR. Using lentiviral technology we provided evidence that it is possible to rescue the dystrophic aberrant phenotype by miRNA669overexpression without gene correction. However, how the viruses carrying the miRNAs were positioned in the genome upon transduction and how their localization site could influence the rescue potential was not analysed. Here we investigate the integration profile of lentiviral vector carrying the pre-miR669 in infected polyclonal and clonal populations derived from Sgcb cardiac progenitors. Our study reveals that the retroviral insertion sites (RIS) are largely restricted to coding genes (65%). Although with the limitation of our analysis, we found no hits for cancer-related genes and several sequenced RIS brought to light genes mainly involved in muscle function. Thus our data show that lentiviral vector insertional profile is cell-specific, however, the chromatin state of target cells positively influences the viral integrations.

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