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EMBRYONIC STEM CELLS/INDUCED PLURIPOTENT STEM CELLS

Herpes Simplex Virus-Mediated Expression of Pax3 and MyoD in Embryoid Bodies Results in Lineage-Related Alterations in Gene Expression Profiles

Departments of ^aMicrobiology and Molecular Genetics and
^bPathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA;
^cDepartment of Bioengineering, University of Pittsburgh School of Engineering, Pittsburgh, Pennsylvania, USA;
^dCogenics, a Division of Clinical Data, Inc., Morrisville, North Carolina, USA;
^eDepartment of Human Genetics, University of Pittsburgh Graduate School of Public Health, Pittsburgh, Pennsylvania, USA

Key Words. Herpes simplex virus • Gene delivery systems in vivo or in vitro • Gene transfer • Embryonic stem cells • Embryoid body • Microarray

Correspondence: Correspondence: Joseph C. Glorioso, Ph.D., E1240 Biomedical Science Tower, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, Pennsylvania 15261, USA. Telephone: 412-648-8106; Fax: 412-624-8997; e-mail: glorioso{at}pitt.edu

Received on May 2, 2008; accepted for publication on August 28, 2008.

First published online in STEM CELLS EXPRESS  September 11, 2008.

The ability of embryonic stem cells to develop into multiple cell lineages provides a powerful resource for tissue repair and regeneration. Gene transfer offers a means to dissect the complex events in lineage determination but is limited by current delivery systems. We designed a high-efficiency replication-defective herpes simplex virus gene transfer vector (JDββ) for robust and transient expression of the transcription factors Pax3 and MyoD, which are known to be involved in skeletal muscle differentiation. JDββ-mediated expression of each gene in day 4 embryoid bodies (early-stage mesoderm) resulted in the induction of unique alterations in gene expression profiles, including the upregulation of known target genes relevant to muscle and neural crest development, whereas a control enhanced green fluorescent protein expression vector was relatively inert. This vector delivery system holds great promise for the use of gene transfer to analyze the impact of specific genes on both regulatory genetic events and commitment of stem cells to particular lineages.

Disclosure of potential conflicts of interest is found at the end of this article.