Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping.
Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping.
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DOI：
10.1534/genetics.115.179648
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发表时间：
2015-09
期刊：
影响因子：
3.3
通讯作者：
Yanik MF
中科院分区：
文献类型：
--
作者：
Gilleland CL;Falls AT;Noraky J;Heiman MG;Yanik MF
人类疾病研究的一个主要目标是为基因或遗传变异分配功能。模式生物秀丽隐杆线虫提供了一个强大的工具，因为许多人类基因的同源物是可识别的，并且大量的遗传载体和突变株是可用的。然而，将这种载体文库递送到突变菌株中仍然是表型分析的长期实验瓶颈。在这里，我们提出了一个计算机辅助显微注射平台，以简化生产转基因C。elegans与多个载体进行深度表型分析。简言之，使用标准多孔平台将动物固定在温度敏感性水凝胶中。然后在自动显微镜的控制下使用由定制的计算机视觉算法驱动的精密机器人进行显微注射。我们证明了效用，通过表型的形态学的12个神经元类在6个突变体的背景下，使用组合的神经元类型特异性的荧光报告。该技术可以通过实现大规模的转基因工程而使体内基因功能的分配工业化。
A major goal in the study of human diseases is to assign functions to genes or genetic variants. The model organism Caenorhabditis elegans provides a powerful tool because homologs of many human genes are identifiable, and large collections of genetic vectors and mutant strains are available. However, the delivery of such vector libraries into mutant strains remains a long-standing experimental bottleneck for phenotypic analysis. Here, we present a computer-assisted microinjection platform to streamline the production of transgenic C. elegans with multiple vectors for deep phenotyping. Briefly, animals are immobilized in a temperature-sensitive hydrogel using a standard multiwell platform. Microinjections are then performed under control of an automated microscope using precision robotics driven by customized computer vision algorithms. We demonstrate utility by phenotyping the morphology of 12 neuronal classes in six mutant backgrounds using combinations of neuron-type-specific fluorescent reporters. This technology can industrialize the assignment of in vivo gene function by enabling large-scale transgenic engineering.