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用于活体动物模型系统中单细胞和多细胞选择性消融的精密光遗传学工具。

Precision Optogenetic Tool for Selective Single- and Multiple-Cell Ablation in a Live Animal Model System.

机构信息

Department of Pharmaceutical Chemistry, University of California - San Francisco, San Francisco, CA 94158, USA; Cardiovascular Research Institute, University of California - San Francisco, San Francisco, CA 94158, USA.

Department of Pharmaceutical Chemistry, University of California - San Francisco, San Francisco, CA 94158, USA; Cardiovascular Research Institute, University of California - San Francisco, San Francisco, CA 94158, USA; Institut Químic de Sarrià, Universitat Ramon Llull, Via Augusta 390, Barcelona 08017, Spain.

出版信息

Cell Chem Biol. 2017 Jan 19;24(1):110-119. doi: 10.1016/j.chembiol.2016.12.010. Epub 2017 Jan 5.

Abstract

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 in live animals, thus opening a new window into cell origin, fate and function, tissue regeneration, and developmental biology.

摘要

细胞消融是一种在发育过程中研究细胞谱系和功能的策略。光遗传学方法是一种重要的细胞消融方法,我们之前开发了一种在活体秀丽隐杆线虫中起作用的迷你单线态氧发生器(miniSOG)工具。在这里,我们通过定向进化生成了 miniSOG2,这是一种通过光生成的活性氧进行细胞消融的改良工具。我们将 miniSOG2 应用于一个更为复杂的模式动物系统——黑腹果蝇,并证明它可用于杀死果蝇幼虫中的单个神经元。此外,miniSOG2 能够在幼虫的一个翅 imaginal 盘上光消融一小群细胞,导致成虫只有一个不完整的和一个正常的翅膀。我们预计 miniSOG2 将成为活体动物中在所需发育时间点进行精确细胞消融的有用光遗传学工具,从而为细胞起源、命运和功能、组织再生和发育生物学开辟新的窗口。

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