Herpes simplex virus-mediated expression of Pax3 and MyoD in embryoid bodies results in lineage-Related alterations in gene expression profiles.
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 (JDbetabeta) for robust and transient expression of the transcription factors Pax3 and MyoD, which are known to be involved in skeletal muscle differentiation. JDbetabeta-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.
Craft
April M
AM
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Krisky
David M
DM
Wechuck
James B
JB
Lobenhofer
Edward K
EK
Jiang
Ying
Y
Wolfe
Darren P
DP
Glorioso
Joseph C
JC
D000818:Animals; D045744:Cell Line, Tumor; D019070:Cell Lineage; D002522:Chlorocebus aethiops; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000821:virology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D018482:Muscle, Skeletal / Q000378:metabolism; D017570:MyoD Protein / Q000096:biosynthesis*; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000096:biosynthesis*; D018139:Simplexvirus / Q000378:metabolism*; D014709:Vero Cells
D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D049452:Green Fluorescent Proteins