Generation of human embryonic stem cells carrying lineage specific reporters.
Generation of human embryonic stem cells carrying lineage specific reporters.
复制标题
产生携带谱系特异性报告基因的人类胚胎干细胞。
作者：
Parinya Noisa;Alai Urrutikoetxea;W. Cui
人类胚胎干细胞（hESC）的独特特性使其能够提供独特的模型来研究调节器官发生的信号通路网络，生成疾病模型，产生用于治疗的细胞和组织，并确定用于治疗的新药。人胚胎干细胞的遗传修饰是辅助上述研究的有力工具。谱系特异性荧光蛋白报告基因hESC系的产生将极大地有利于研究者以实时、简单和及时的方式监测特定细胞谱系。该技术将促进高通量筛选以鉴定在调节特定细胞命运定型中重要的分子。此外，这种报告细胞系使研究人员能够通过荧光激活细胞分选（FACS）富集某些细胞群，用于下游生物分析或体内应用。我们已经表明，hESC可以稳定转染的质粒中的绿色荧光蛋白（GFP）的表达是在控制的Oct-4启动子使用化学转染。转基因Oct-4-GFP的表达模式反映了内源Oct-4的表达模式。
The distinctive properties of human embryonic stem cells (hESCs) enable them to provide unique models to study the network of signaling pathways that regulate organogenesis, generate disease models, produce cells and tissues for therapies, and identify new drugs for treatment. Genetic modification of hESCs is a powerful tool to assist the above studies. Generation of lineage-specific fluorescent protein reporter hESC lines will greatly benefit investigators to monitor specific cell lineages in a live, easy, and timely manner. This technique will facilitate high throughput screening to identify molecules important in regulating specific cell fate commitment. In addition, such reporter cell lines enable researchers to enrich certain cell populations by fluorescent activated cell sorting (FACS) for either downstream biological analysis or in vivo applications. We have shown that hESCs can be stably transfected with a plasmid in which expression of the green fluorescent protein (GFP) is under the control of the Oct-4 promoter using chemical transfection. The expression pattern of transgenic Oct-4-GFP reflects that of endogenous Oct-4.