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Reporter Genes02:11

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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.
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Ribosome Profiling02:24

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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.
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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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自己報告型トランポゾンは,単細胞における遺伝子発現と転写因子の結合を同時に読み出すことを可能にします.

Arnav Moudgil1, Michael N Wilkinson2, Xuhua Chen2

  • 1Department of Genetics, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Edison Family Center for Genome Sciences and Systems Biology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA; Medical Scientist Training Program, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.

Cell
|July 26, 2020
PubMed
まとめ
この要約は機械生成です。

単細胞通話カード (scCC) を開発して 個々の細胞の転写因子結合部位 (TFBS) をマッピングしました この新しい方法は,細胞のアイデンティティとTFBSを結びつけ,複雑な組織における遺伝子調節の研究を進めています.

キーワード:
ブロモドメイン電話カード細胞状態マウス皮質シングルセル転写因子トランポゾン

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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:

  • 細胞の異質性は,転写因子 (TF) 結合の局所分析を複雑にする.
  • 現在の単細胞RNAシーケンシング (scRNA-seq) 方法は,細胞タイプを特定できますが,特定のTF結合部位 (TFBS) との細胞同一性をリンクすることはできません.

研究 の 目的:

  • 単細胞でのTFBSの遺伝子発現とマッピングを同時に行う新しい測定法を開発する.
  • 細胞のアイデンティティとTFの結合を in situ 関連付ける方法を確立する.

主な方法:

  • mRNAから回収できる自己報告トランポゾン (SRT) の開発.
  • TFBSをマッピングするために,SRTとTF-トランスポゼ融合を統合する.
  • scRNA-seqとSRTマッピングを組み合わせた単細胞呼び出しカード (scCC) 測定の適用,同時に遺伝子発現とTFBS分析を行う.

主要な成果:

  • scCCの複数のTFとのベンチマークは,その有効性を示した.
  • scCCを用いたK562細胞におけるBRD4媒介細胞状態移行の発見.
  • 単細胞解像度でのマウス皮質のBRD4結合部位のマッピング.

結論:

  • 単細胞電話カード (scCC) は,TF生物学を現地で研究するための強力な新しい方法を提供します.
  • scCC測定法により,細胞タイプとその関連TFBSを同時に特定できます.
  • この技術は単細胞レベルで 遺伝子調節と細胞状態のダイナミクスを理解するための 新しい道を開きます