相关实验视频
Updated: Jun 13, 2025
11:03
Massively Parallel Reporter Assays in Cultured Mammalian Cells
Published on: August 17, 2014
21.5K
在单核酸分辨率下测绘基因特异性调节区域的大规模并行记者试验
Alastair J Tulloch1,2, Ryan N Delgado1,2, Rinaldo Catta-Preta1,2,3
1Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
bioRxiv : the preprint server for biology
|June 4, 2025
概括
新的特定位置 (LS-MPRA) 和退化的 (d-MPRA) 大规模并行记者测定有效地映射特定细胞类型的 cis-regulatory 模块 (CRMs) 和转录因子结合位.
科学领域:
- 基因组学和分子生物学
- 发展生物学 发展生物学
- 神经科学是一个神经科学.
背景情况:
- 精确的基因调节对于组织发育和功能至关重要.
- 目前的技术限制了 cis 监管模块 (CRM) 的全面映射.
- 现有的方法缺乏吞吐量,分辨率和细胞类型特异性.
研究的目的:
- 引入用于高分辨率 cis 调节模块发现的新型,互补的方法.
- 为了能够在大型基因组区域中对监管要素进行公正的审问.
- 为特定的基因在CRMs中识别转录因子结合部位.
主要方法:
- 使用细菌人工染色体 (BACs) 的局部特异性大规模并行记者测定 (LS-MPRA).
- 退化MPRA (d-MPRA) 采用系统性突变生成来实现核酸水平的分辨率.
- 适用于小鼠,小视网膜和脊髓组织中的视网膜基因.
主要成果:
- LS-MPRA成功地确定了视网膜基因的已知和新型CRMs,包括邻近基因中的基因.
- 对于动态表达的基因Olig2.2,LS-MPRA确定了三个不同的CRM区域.
- d-MPRA和基因分析提名了关键转录因子结合位点,由CUT&RUN实验证实.
结论:
- 集成的LS-MPRA和d-MPRA提供了一个强大的,高分辨率的平台,用于cis-regulatory代码发现.
- 这些方法可以在没有事先知识的情况下快速,可扩展和负担得起的CRM发现.
- 该平台适用于各种物种和组织,促进细胞类型特定的基因表达研究.
更多相关视频
10:46Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
Published on: June 2, 2018
9.3K
11:36Author Spotlight: An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations
Published on: April 21, 2023
2.0K
相关概念视频
02:11Reporter Genes
11.3K
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.
11.3K
02:34DNA Microarrays
17.2K
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
17.2K
02:24Ribosome Profiling
3.5K
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...
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...
3.5K