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関連する概念動画

RNA Editing02:23

RNA Editing

9.2K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Leaky Scanning02:28

Leaky Scanning

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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...
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From DNA to Protein03:06

From DNA to Protein

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The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
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The Central Dogma01:25

The Central Dogma

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Overview
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tRNA Activation02:26

tRNA Activation

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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

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The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
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Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
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プログラム可能な偽ウリジン編集と解読によるRNAコドン拡張

Jiangle Liu1,2,3, Xueqing Yan1, Hao Wu1,3

  • 1The National Key Laboratory of Gene Function Studies and Manipulation, School of Life Sciences, Peking University, Beijing, China.

Nature
|June 25, 2025
PubMed
まとめ
この要約は機械生成です。

研究者は,哺乳類の細胞に非正規アミノ酸 (ncAA) を正確に組み込むために,偽ウリジン (Ψ) コドンを使用して,新しいRNAコドン拡張 (RCE) 戦略を開発した. この方法は特異性を高め 遺伝子アルファベットの拡張のための新しい経路を提供します

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers
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関連する実験動画

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Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
11:47

Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers
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Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers

Published on: June 24, 2019

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科学分野:

  • 合成生物学
  • 分子生物学
  • 生物化学

背景:

  • 非正規のアミノ酸 (ncAAs) は,カスタム化学を通じて特別のタンパク質機能を可能にします.
  • 遺伝子コード拡張 (GCE) は,ncAA組み込みのためのストップコドン再割り当てを使用するが,完全な正交性がない.
  • セルラーシステムにおけるncAAエンコーディングのためのより具体的で直角な方法が必要である.

研究 の 目的:

  • サイト固有のncAAの組み込みのための新しいRNAコドン拡張 (RCE) 戦略を開発する.
  • 翻訳特異性を高めるために,偽ウリジン (Ψ) を使用してバイオオルトゴナル"空白"コドンを作成する.
  • ユカリオット細胞の遺伝子アルファベットを拡張する方法を確立する.

主な方法:

  • プログラム可能なガイドRNA,エンジニアリングされたtRNA,およびアミノアシル-tRNA合成酵素を含むRCE戦略を開発した.
  • 特定のmRNAトランスクリプトに Pseudouridine (Ψ) コドン (ΨGA, ΨAA, ΨAG) を導入し,解読した.
  • 哺乳類の細胞におけるRCEシステムの正交性と特異性を試験した.

主要な成果:

  • RCE (ΨGA) システムは,GCEよりも高いトランスラトーム全体およびタンパク質特異性を示した.
  • 確立されたRCE (ΨAA) とRCE (ΨAG) システムで,3つの Ψ コドンペアの相互正交を示している.
  • RCEとGCEの間の互換性のある協力が証明されている.

結論:

  • RCE戦略は,特定のRNAコドンをコーディングするためのポストトランスクリプション要素としてPzを効果的に使用します.
  • この方法は,真核細胞におけるサイト固有のncAAの組み込みのための新しい,高度に特異的な経路を提供します.
  • RCEは遺伝子アルファベットの拡張とカスタムタンパク質工学の可能性を広げています.