相关实验视频

Updated: Jul 11, 2025

Massively Parallel Reporter Assays in Cultured Mammalian Cells

Massively Parallel Reporter Assays in Cultured Mammalian Cells

Published on: August 17, 2014

一个依赖于Cre的大规模并行导体试验允许对细胞类型进行特定的 in vivo 非编码元素的功能影响的评估

Tomas Lagunas^1,2,3, Stephen P Plassmeyer^1,2, Anthony D Fischer^1,2

¹Department of Genetics, Washington University School of Medicine, 660 S. Euclid Ave, Saint Louis, MO, 63108, USA.

Communications biology

|November 12, 2023

概括

研究人员开发了一种新方法,在体内测试特定脑细胞中的基因调节元件. 这种技术有助于通过研究神经元中的元素来理解与自闭症等精神疾病的遗传联系.

科学领域:

神经科学是一个神经科学.
遗传学是一个遗传学.
分子生物学分子生物学

背景情况:

调控元件的功能是特定于细胞类型的,这对于精神疾病研究至关重要.
现有的大规模并行报告测试 (MPRA) 在复杂组织中缺乏体内细胞类型的特异性.
研究与精神疾病相关的因素需要体内神经元背景.

研究的目的:

为在小鼠大脑中进行细胞类型特定的体内分析而调整大规模并行报告测试 (MPRA).
为了能够在特定的神经元群体内对调节元件进行功能查.
为了促进与精神疾病相关的遗传变异在它们的原生细胞环境中的研究.

主要方法:

开发了一种新的MPRA技术,用于在小鼠大脑神经元中进行有针对性的体内查.
使用一个测试案例3'UTR MPRA库与自闭症患者衍生的变体.
在激发性皮质神经元和条状中等脊状神经元中验证了该方法.

主要成果:

在体内实现了可重复的,细胞类型特定的调节元件功能的测量.
在复杂的大脑组织中证明了向MPRA的可行性.
成功地应用该方法来研究与自闭症相关的遗传元素.

结论:

开发的向MPRA技术能够在特定的神经元环境中对遗传元素进行强大的功能注释.
这种方法对于理解心理疾病中调节因素的作用至关重要.
促进与神经和精神疾病相关的细胞类型的体内功能基因组学.

更多相关视频

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Published on: June 2, 2018

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

Published on: March 31, 2022

相关概念视频

Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.

Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...