Nanobody L10 Method
- 详细技术说明
- There exists a critical, unmet need to molecularly profile defined cell-types throughout the mammalian brain. A neuron can be defined in multiple ways. For example, principal genetic markers, activity, and connectivity can be used to identify a cell type. With the advent of GENSAT, GFP reporter lines exist for a broad array of genetically-defined cell types throughout the brain, but there is no way to comprehensively profile these neurons. The current work describes a methodology for capturing the molecular profile of any cell type expressing GFP.
- *Abstract
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Our scientists have created a unique transgenic mouse strain that expresses an anti-GFP nanobody fused to the ribosomal subunit protein Rpl10a (Nanobody-L10). When this mouse is used in tandem with any GFP expression system, it enables the selective capture of translating mRNAs from the cell types expressing GFP. For example, this mouse can be used in tandem with any tracing virus expressing GFP for molecular profiling of neurons based on connectivity. Additionally, this mouse can be crossed with another mouse that expresses a promoter of interest driving GFP to purify mRNAs from that cell type. The resulting complex containing GFP/nanobody/ribosome can easily be purified. This system allows for the identification and molecular profiling of cell populations expressing GFP.
- 国家/地区
- 美国
