通过生物基因组编辑对哺乳动物大脑中的蛋白质定位进行高通量,高分辨率的映射
Takayasu Mikuni1, Jun Nishiyama1, Ye Sun2
1Neuronal Signal Transduction Group, Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA.
Cell
|May 17, 2016
概括
我们开发了一种基于CRISPR的方法, 这种高通量技术为了解细胞功能提供纳米分辨率.
科学领域:
- 神经科学
- 分子生物学
- 细胞生物学
背景情况:
- 了解蛋白质局部化对于细胞生物学来说至关重要.
- 现有的方法缺乏内源蛋白质的可扩展性和精度.
研究的目的:
- 开发一种高通量,高分辨率的成像方法,用于在单个哺乳动物脑细胞中定位内源蛋白.
主要方法:
- 通过CRISPR-Cas9介导的同质导向修复 (SLENDR) 开发了内源蛋白的单细胞标记.
- 通过CRISPR-Cas9和同源导向修复 (HDR) 使用体内基因组编辑.
- 通过在子宫内进行单细胞基因组编辑,向神经元前代传递编辑机器.
主要成果:
- SLENDR能够快速确定许多内源蛋白的局部和动态.
- 在单个脑细胞中表现出高的特异性,分辨率 (微米至纳米) 和对比度.
- 已成功应用于大脑中的不同细胞类型,区域和年龄.
结论:
- SLENDR是一种可通用和可扩展的技术,用于绘制细胞下蛋白质定位.
- 提供了对细胞分子功能的综合理解的强大平台.
- 便于对大脑中的蛋白质动态进行高通量分析.
更多相关视频
相关概念视频
01:26Cerebrum: Anatomical Overview I
The main and largest component of the human brain is the cerebrum. The cerebrum consists of two main parts: the cerebral cortex, an outer layer with wrinkles or folds known as gyri and shallow grooves called sulci, and a deeper region beneath it. The cerebrum divides into two distinct hemispheres and contains five different lobes: the frontal, parietal, temporal, occipital, and insula. The central sulcus separates the frontal and parietal lobes and two functionally important gyri — the...
01:11Cerebrum: Anatomical Overview II
Each cerebral hemisphere can be divided into three main regions. The outermost region, the cerebral cortex, is a thin layer (2 to 4 millimeters thick) made up of gray matter, consisting of neuron cell bodies, dendrites, glial cells, and blood vessels. The middle region, or white matter, is primarily composed of myelinated nerve fibers organized into three types of large tracts: association fibers, commissures, and projection fibers. Association fibers connect different areas within the same...