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RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Ribosome Profiling02:24

Ribosome Profiling

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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...
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RNA Stability01:53

RNA Stability

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Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
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Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
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単細胞における代謝的に標識されたトランスクリプトの配列化は,mRNAのターンオーバー戦略を明らかにする.

Nico Battich1, Joep Beumer2, Buys de Barbanson2

  • 1Oncode Institute, Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences) and University Medical Center Utrecht, 3584 CT Utrecht, Netherlands. n.battich@hubrecht.eu a.vanoudenaarden@hubrecht.eu.

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|March 7, 2020
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まとめ

研究者は単細胞でのメッセンジャーRNAの合成と分解率を測定する新しい方法を開発しました. このテクニックは 細胞が遺伝子発現を 微分化や細胞サイクルのような ダイナミックなプロセスで制御する方法を示しています

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科学分野:

  • 分子生物学
  • ゲノミクス
  • 細胞生物学

背景:

  • 哺乳類の細胞におけるメッセンジャーRNA (mRNA) レベルは,合成と分解率によって調節される.
  • 以前の大量RNA標識法では,複雑な細胞プロセスに必要な単細胞解像度が欠けていました.
  • 異質な細胞集団における遺伝子発現のダイナミクスを理解することは極めて重要です.

研究 の 目的:

  • 単細胞レベルでラベルとラベルのないトランスクリプトを同時に定量化するための方法を開発する.
  • 細胞サイクルと腸の幹細胞の分化中にmRNA合成と分解速度を決定する.
  • 遺伝子発現のダイナミクスを制御する規制戦略を特定する.

主な方法:

  • 新しく合成されたと既存のトランスクリプトの同時定量化を可能にする新しい代謝RNAラベル技術を開発した.
  • 遺伝子発現のダイナミクスを分析するために 何千もの個別の細胞に適用しました
  • 細胞サイクルの進行と腸の幹細胞の分化における研究率

主要な成果:

  • 異なる細胞状態における個々の細胞におけるmRNAの量化合成と分解率.
  • 細胞が細胞サイクルと分化過程で用いる異なる制御戦略を特定した.
  • これらの戦略が遺伝子発現のダイナミックレンジと精度にどのように影響するかを示した.

結論:

  • 開発された方法は,mRNAのダイナミクスを研究するための前例のない単細胞解像度を提供します.
  • 細胞の分化と細胞サイクルには,遺伝子発現のための特定の規制戦略が含まれています.
  • この発見により,異質な細胞集団における遺伝子発現制御の理解が進んでいます.