Craft April M, Krisky David M, Wechuck James B, Lobenhofer Edward K, Jiang Ying, Wolfe Darren P, Glorioso Joseph C
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Stem Cells. 2008 Dec;26(12):3119-29. doi: 10.1634/stemcells.2008-0417. Epub 2008 Sep 11.
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.
胚胎干细胞发育成多种细胞谱系的能力为组织修复和再生提供了强大的资源。基因转移为剖析谱系决定中的复杂事件提供了一种手段，但受到当前递送系统的限制。我们设计了一种高效复制缺陷型单纯疱疹病毒基因转移载体（JDbetabeta），用于转录因子Pax3和MyoD的强大且瞬时表达，已知这两种转录因子参与骨骼肌分化。JDbetabeta介导的每个基因在第4天胚状体（早期中胚层）中的表达导致基因表达谱中独特变化的诱导，包括与肌肉和神经嵴发育相关的已知靶基因的上调，而对照增强型绿色荧光蛋白表达载体相对无活性。这种载体递送系统在利用基因转移分析特定基因对调控遗传事件以及干细胞向特定谱系定向分化的影响方面具有巨大潜力。
