Precision Optogenetic Tool for Selective Single- and Multiple-Cell Ablation in a Live Animal Model System.
Precision Optogenetic Tool for Selective Single- and Multiple-Cell Ablation in a Live Animal Model System.
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DOI：
10.1016/j.chembiol.2016.12.010
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发表时间：
2017-01-19
影响因子：
8.6
通讯作者：
Shu X
中科院分区：
文献类型：
--
作者：
Makhijani K;To TL;Ruiz-González R;Lafaye C;Royant A;Shu X
细胞消融是研究发育过程中细胞谱系和功能的一种策略。光遗传学方法是一种重要的细胞消融方法，我们以前已经开发了一种小型单线态氧发生器（miniSOG）工具，可以在活体秀丽隐杆线虫中工作。在这里，我们使用定向进化来生成miniSOG 2，这是一种通过光生活性氧进行细胞消融的改进工具。我们将miniSOG 2应用于更复杂的模型动物系统，果蝇，并证明它可以用来杀死果蝇幼虫中的单个神经元。此外，miniSOG 2能够光消融一个幼虫翅盘中的一小群细胞，导致一个成虫具有一个不完整的翅和一个正常的翅。我们期望miniSOG 2成为一种有用的光遗传学工具，用于在活体动物中所需的发育时间点进行精确的细胞消融，从而为细胞起源，命运和功能，组织再生和发育生物学打开新的窗口。
Cell ablation is a strategy to study cell lineage and function during development. Optogenetic methods are an important cell-ablation approach, and we have previously developed a mini singlet oxygen generator (miniSOG) tool that works in the living Caenorhabditis elegans. Here, we use directed evolution to generate miniSOG2, an improved tool for cell ablation via photogenerated reactive oxygen species. We apply miniSOG2 to a far more complex model animal system, Drosophila melanogaster, and demonstrate that it can be used to kill a single neuron in a Drosophila larva. In addition, miniSOG2 is able to photoablate a small group of cells in one of the larval wing imaginal discs, resulting in an adult with one incomplete and one normal wing. We expect miniSOG2 to be a useful optogenetic tool for precision cell ablation at a desired developmental time point inlive animals, thus openinga new window into cell origin, fate and function, tissue regeneration, and developmental biology.