Chan Yick-Bun, Alekseyenko Olga V, Kravitz Edward A
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, United States of America.
PLoS One. 2015 Sep 18;10(9):e0138181. doi: 10.1371/journal.pone.0138181. eCollection 2015.
To study the molecular mechanism of complex biological systems, it is important to be able to artificially manipulate gene expression in desired target sites with high precision. Based on the light dependent binding of cryptochrome 2 and a cryptochrome interacting bHLH protein, we developed a split lexA transcriptional activation system for use in Drosophila that allows regulation of gene expression in vivo using blue light or two-photon excitation. We show that this system offers high spatiotemporal resolution by inducing gene expression in tissues at various developmental stages. In combination with two-photon excitation, gene expression can be manipulated at precise sites in embryos, potentially offering an important tool with which to examine developmental processes.
为了研究复杂生物系统的分子机制,能够在所需靶位点高精度地人工操纵基因表达非常重要。基于隐花色素2与隐花色素相互作用的bHLH蛋白的光依赖性结合,我们开发了一种用于果蝇的分裂LexA转录激活系统,该系统允许使用蓝光或双光子激发在体内调节基因表达。我们表明,该系统通过在不同发育阶段的组织中诱导基因表达,提供了高时空分辨率。与双光子激发相结合,可以在胚胎的精确位点操纵基因表达,这可能为研究发育过程提供一个重要工具。