Takagi, Shin et al. (2009) International Worm Meeting "IR-LEGO: a tool for manipulating gene expression in targeted single cells in vivo ."

Suzuki, Motoshi, Takagi, Shin, Yuba, Shunsuke, Kamei, Yasuhiro

[International Worm Meeting, 2009]

We have developed a novel microscope system, Infrared Laser Evoked Gene Operator (IR-LEGO), for inducing gene expression in vivo. By depositing heat under the microscope with irradiation of infrared laser, IR-LEGO induces the heat shock response in targeted single cells, thereby eliciting the expression of a gene of interest under the control of a heat shock promoter in transgenes. Using C. elegans as an experimental material, we have been testing applicability of IR-LEGO to living organisms, and determined the optimum condition for gene induction. A comparison with a conventional visible laser (440-nm dye laser) system for cell ablation experiments showed that the IR-laser (1480 nm) is much more suitable for inducing gene expression; With the IR-laser, gene induction can be induced in larval seam cells more efficiently under a wider range of conditions, with an irradiation period of less than one second. While the visible laser often has detrimental effects on irradiated cells, the IR-laser is far less harmful. Thus, IR-LEGO would be a useful tool for C. elegans research, particularly in analyzing development as well as manipulating the function of nervous system. Future prospects as well as current technical limitations of this method will be discussed.