Wang Yisen, He Hao, Li Shiyang, Liu Dayong, Lan Bei, Hu Minglie, Cao Youjia, Wang Chingyue
Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin, 300072, P.R. China.
1] Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin, 300072, P.R. China [2] Med-X Research Institute, School of Biomedical Engineering, Shanghai Jiaotong University, Shanghai, 200030, P.R. China.
Sci Rep. 2014 Jun 18;4:5346. doi: 10.1038/srep05346.
Controllable gene expression is always a challenge and of great significance to biomedical research and clinical applications. Recently, various approaches based on extra-engineered light-sensitive proteins have been developed to provide optogenetic actuators for gene expression. Complicated biomedical techniques including exogenous genes engineering, transfection, and material delivery are needed. Here we present an all-optical method to regulate gene expression in targeted cells. Intrinsic or exogenous genes can be activated by a Ca(2+)-sensitive transcription factor nuclear factor of activated T cells (NFAT) driven by a short flash of femtosecond-laser irradiation. When applied to mesenchymal stem cells, expression of a differentiation regulator Osterix can be activated by this method to potentially induce differentiation of them. A laser-induced "Ca(2+)-comb" (LiCCo) by multi-time laser exposure is further developed to enhance gene expression efficiency. This noninvasive method hence provides an encouraging advance of gene expression regulation, with promising potential of applying in cell biology and stem-cell science.
可控基因表达一直是一项挑战，对生物医学研究和临床应用具有重要意义。最近，基于额外工程化的光敏感蛋白开发了各种方法，以提供用于基因表达的光遗传学激活器。这需要包括外源基因工程、转染和材料递送在内的复杂生物医学技术。在此，我们提出一种全光学方法来调节靶细胞中的基因表达。内在基因或外源基因可由飞秒激光短脉冲照射驱动的钙敏感转录因子活化T细胞核因子（NFAT）激活。当应用于间充质干细胞时，这种方法可激活分化调节因子Osterix的表达，从而有可能诱导它们分化。通过多次激光照射进一步开发了激光诱导的“钙梳”（LiCCo）以提高基因表达效率。因此，这种非侵入性方法为基因表达调控带来了令人鼓舞的进展，在细胞生物学和干细胞科学中具有广阔的应用潜力。
