A Mammalian enhancer trap resource for discovering and manipulating neuronal cell types.
A Mammalian enhancer trap resource for discovering and manipulating neuronal cell types.
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DOI：
10.7554/elife.13503
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发表时间：
2016-03-21
期刊：
影响因子：
7.7
通讯作者：
Nelson SB
中科院分区：
文献类型：
--
作者：
Shima Y;Sugino K;Hempel CM;Shima M;Taneja P;Bullis JB;Mehta S;Lois C;Nelson SB
为了在神经系统中实现细胞类型特异性操作，仍然需要驱动菌株。每种细胞类型表达一组独特的基因，通过BAC转基因或敲入对标记基因的重述表达产生了有用的转基因小鼠系。然而，由于基因通常在许多细胞类型中表达，许多这些细胞系具有相对广泛的表达模式。我们报告了一种替代的转基因方法，捕获远端增强子以获得更集中的表达。我们发现了一种增强子陷阱探针，经常产生受限的报告基因表达，并利用PiggyBac转座子开发了有效的增强子陷阱筛选。我们建立了200多个细胞系，并发现许多细胞系标记了大脑亚结构中的小子集神经元，包括已知的和新的细胞类型。每条生产线的图片和其他信息都可以在网上找到（enhancertrap.bio.branddeis.edu）。DOI: http://dx.doi.org/10.7554/eLife.13503.001科学家可以通过操纵神经元的基因来追踪甚至改变不同种类神经元的活动，以及神经元之间的联系。然而，大多数基因在大脑中许多不同位置的许多不同种类的细胞中都是活跃的，因此很难追踪或仅针对特定的神经元或大脑区域。增强子是DNA的一部分，它可以调节附近基因的活动，使它们只在非常特定的细胞类型中活跃，而“增强子陷阱”是一种基因方法，本质上劫持增强子，在相同的细胞类型中表达人工基因。这项技术依赖于在基因组的随机位置插入一个易于追踪的遗传标记。如果这个标记与一个增强子相互作用，它就会被激活，增强子对基因表达的影响就可以被评估。这种方法已在果蝇和鱼类中用于鉴定特异性地将基因表达限制在一小部分细胞中的增强子。现在，Shima等人表明，增强诱捕器也可以成功地用于哺乳动物。实验产生了200多种不同品系的老鼠，其中许多只在特定的大脑区域或特定种类的脑细胞中有荧光标记。这些实验发现的一些脑细胞类型是新的，在不同品系中标记特定的脑细胞和大脑区域使这些小鼠成为未来工作的有用资源。此外，生产更多的这些小鼠品系将是相对简单的，因为它只涉及杂交小鼠。其中一些待发现的菌株很可能也会成为研究的有用工具。DOI: http://dx.doi.org/10.7554/eLife.13503.002
There is a continuing need for driver strains to enable cell-type-specific manipulation in the nervous system. Each cell type expresses a unique set of genes, and recapitulating expression of marker genes by BAC transgenesis or knock-in has generated useful transgenic mouse lines. However, since genes are often expressed in many cell types, many of these lines have relatively broad expression patterns. We report an alternative transgenic approach capturing distal enhancers for more focused expression. We identified an enhancer trap probe often producing restricted reporter expression and developed efficient enhancer trap screening with the PiggyBac transposon. We established more than 200 lines and found many lines that label small subsets of neurons in brain substructures, including known and novel cell types. Images and other information about each line are available online (enhancertrap.bio.brandeis.edu). DOI: http://dx.doi.org/10.7554/eLife.13503.001 Scientists can track and even alter the activity of different kinds of neurons, as well as the connections between neurons, by manipulating their genes. However, most genes are active in many different kinds of cells in many different places in the brain, making it difficult to track or target only a particular neuron or brain area. Enhancers are sections of DNA that can regulate the activity of nearby genes so that they are only active in very specific cell types, and an “enhancer trap” is a genetic approach that essentially hijacks enhancers to express artificial genes in those same cell types. The technique relies on inserting a genetic marker, which can be easily tracked, into random locations in the genome. If this marker then interacts with an enhancer, it is activated and the effect of the enhancer on gene expression can be assessed. This method has been used in fruit flies and fish to identify enhancers that specifically restrict gene expression to a small subset of cells. Now, Shima et al. show that enhancer traps can be used successfully in mammals too. The experiments produced over 200 different strains of mice, many with the fluorescent marker only in specific brain areas or in specific kinds of brain cells. Some of the types of brain cells uncovered by these experiments are new, and the labelling of specific brain cells and brain areas in different strains makes these mice a useful resource for future work. Furthermore, it will be relatively straightforward to produce many more strains of these mice, because it would simply involve crossbreeding mice. It is likely that some of these to-be-discovered strains will be useful tools for research as well. DOI: http://dx.doi.org/10.7554/eLife.13503.002