Fast targeted gene transfection and optogenetic modification of single neurons using femtosecond laser irradiation.
Fast targeted gene transfection and optogenetic modification of single neurons using femtosecond laser irradiation.
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DOI：
10.1038/srep03281
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发表时间：
2013-11-21
影响因子：
4.6
通讯作者：
Gunn-Moore, Frank J.
中科院分区：
文献类型：
--
作者：
Antkowiak, Maciej;Torres-Mapa, Maria Leilani;Witts, Emily C.;Miles, Gareth B.;Dholakia, Kishan;Gunn-Moore, Frank J.
神经科学中的一个普遍问题是将DNA快速和有针对性地递送到选定的神经元中。适当方法学的发展将使得能够将多个基因转染到同一细胞中或将不同基因转染到不同的邻近细胞中以及神经元的快速细胞选择性功能化。在这里，我们表明，优化的飞秒光学转染满足这些要求。我们还证明了成功的光学转染channelrhodopsin-2在单个选定的神经元。我们扩展了这项技术的功能，通过使用快速的三维激光束转向，使图像引导的“点和扫描”用户友好的转染选定的细胞更广泛的吸收神经科学家。亚秒级的转染时间尺度/细胞使得该方法与替代的单细胞转染技术相比更快至少两个数量级。这项新技术提供了对神经元集合进行大规模细胞选择性遗传研究和对神经回路进行快速遗传编程的能力。
A prevailing problem in neuroscience is the fast and targeted delivery of DNA into selected neurons. The development of an appropriate methodology would enable the transfection of multiple genes into the same cell or different genes into different neighboring cells as well as rapid cell selective functionalization of neurons. Here, we show that optimized femtosecond optical transfection fulfills these requirements. We also demonstrate successful optical transfection of channelrhodopsin-2 in single selected neurons. We extend the functionality of this technique for wider uptake by neuroscientists by using fast three-dimensional laser beam steering enabling an image-guided “point-and-transfect” user-friendly transfection of selected cells. A sub-second transfection timescale per cell makes this method more rapid by at least two orders of magnitude when compared to alternative single-cell transfection techniques. This novel technology provides the ability to carry out large-scale cell selective genetic studies on neuronal ensembles and perform rapid genetic programming of neural circuits.
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影响因子：
48
作者：
Barrett, Lindy E.;Sul, Jai-Yoon;Eberwine, James H.
通讯作者：
Eberwine, James H.
影响因子：
2.8
作者：
Antkowiak, Maciej;Torres-Mapa, Maria Leilani;Dholakia, Kishan
通讯作者：
Dholakia, Kishan
影响因子：
9.2
作者：
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通讯作者：
Gottschalk, A
影响因子：
56.9
作者：
Naldini, L;Blomer, U;Trono, D
通讯作者：
Trono, D
影响因子：
1.7
作者：
Barlow, H B
通讯作者：
Barlow, H B