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相关概念视频

Ribosome Profiling02:24

Ribosome Profiling

3.7K
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...
3.7K
Translational Regulation01:29

Translational Regulation

273
Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
273
Stringent Response in E. coli01:23

Stringent Response in E. coli

92
Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...
92
Leaky Scanning02:28

Leaky Scanning

5.3K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.3K
Improving Translational Accuracy02:07

Improving Translational Accuracy

12.0K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
12.0K
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

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相关实验视频

Updated: Oct 21, 2025

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
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单细胞Ribo-seq显示细胞周期依赖的转化暂停

Michael VanInsberghe1, Jeroen van den Berg2, Amanda Andersson-Rolf2

  • 1Oncode Institute, Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences) and University Medical Center Utrecht, Utrecht, The Netherlands. m.vaninsberghe@hubrecht.eu.

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概括

研究人员开发了一种敏感的单细胞核糖体分析方法. 这种技术与机器学习相结合,实现了编码分辨率以研究翻译和细胞间的变异.

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RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling
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Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling

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相关实验视频

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RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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科学领域:

  • 分子生物学
  • 基因组学
  • 细胞生物学

背景情况:

  • 单细胞测序推进了基因组,表观基因组和转录组分析.
  • 测量单细胞水平的翻译仍然是一个重大挑战.
  • 现有的蛋白质检测方法缺乏单细胞分辨率进行翻译.

研究的目的:

  • 在单个细胞中开发一种高度敏感的核糖体分析方法.
  • 为了实现单个编码分辨率来分析细胞翻译.
  • 研究细胞多样性中的翻译作用.

主要方法:

  • 改进了核糖体分析方案以提高敏感性.
  • 整合机器学习以实现单个编码器的分辨率.
  • 在各种细胞环境中验证,包括罕见的细胞类型.

主要成果:

  • 已证实氨基酸限制导致核糖体在特定的子中暂停.
  • 观察到细胞循环依赖的核糖体暂停.
  • 在转化过程中检测出明显的GAA码头暂停.
  • 这项技术应用于罕见的初级肠内分泌细胞.

结论:

  • 开发的技术可以对单细胞翻译进行敏感,高分辨率的分析.
  • 核糖体暂停与特定的细胞状态有关,如细胞循环和细胞分裂.
  • 这种方法适用于罕见的细胞群,开辟了新的研究途径.
  • 为了解细胞多样性的转化贡献提供了基础工具.