Dougherty Joseph D
Departments of Genetics and Psychiatry, Washington University School of Medicine, St. Louis, Missouri 63110
J Neurosci. 2017 Dec 13;37(50):12079-12087. doi: 10.1523/JNEUROSCI.1929-17.2017.
Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation. These advances include a variety of new methods for targeting cells for translating ribosome affinity purification by features such as their projections or activity, additional tags and mouse reagents increasing the flexibility of the system, and new modifications of the method specifically focused on studying the regulation of translation. The latter includes methods to assess cell type-specific regulation of translation in specific subcellular compartments. Here, I provide a summary of these recent advances and resources, highlighting both new experimental opportunities and areas for future technical development.
翻译核糖体亲和纯化是一种最初开发用于分析复杂组织中特定基因定义细胞类型的mRNA的方法。它已被应用于识别对控制大脑中各种行为很重要的细胞类型中的靶分子,以及理解由于实验操作(从滥用药物到致病突变)而对这些细胞产生的分子后果。自该方法问世以来,各种方法学进展正在开辟新的研究途径。这些进展包括多种通过细胞投射或活性等特征靶向细胞进行翻译核糖体亲和纯化的新方法、增加系统灵活性的额外标签和小鼠试剂,以及专门侧重于研究翻译调控的该方法的新改进。后者包括评估特定亚细胞区室中细胞类型特异性翻译调控的方法。在此,我总结了这些最新进展和资源,突出了新的实验机会和未来技术发展的领域。