アルキニルフォスフォネートDNA:DNAベースの生物学的応用のための多用途の"クリック"可能なバックボーン
Heera Krishna1, Marvin H Caruthers
1Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, Colorado 80309, USA.
Journal of the American Chemical Society
|May 23, 2012
まとめ
研究者は,多用途のトリアゾリルフォスフォナート (TP) DNA アナログを作成するための新しい"クリック"化学方法を開発しました. これらの改変オリゴヌクレオチド (ODN) は,安定性と細胞吸収の改善を示し,生物学的研究のための新しいツールを提供しています.
科学分野:
- オリゴヌクレオチド化学 オリゴヌクレオチド化学
- 化学生物学 化学生物学とは
- バイオテクノロジー バイオテクノロジー
背景:
- DNAベースのアプリケーションは,細胞配送,非特異的な効果,毒性,および酵素安定性に関する課題に直面しています.
- 既存のオリゴヌクレオチド類型は,これらの問題を体系的に解決するための多用途な方法論を欠いている.
研究 の 目的:
- DNAアナログの迅速な機能化のための新しい,クリック可能な化学を導入する.
- トリアゾリルフォスフォナート (TP) 改変オリゴヌクレオチド (ODN) の生物学的性質を合成し評価する.
主な方法:
- 固体相合成と互換性のあるアルキニルフォスフォラミドイトシントンの開発.
- "クリック"化学を用いたポスト合成機能化により,TPの核酸間結合を作り出します.
- 2'-OMeおよびLNA改変を含むTP,フォスファート,またはチオフォスファート結合による混合骨格ODNの自動固相合成.
主要な成果:
- TP結合は,5'-および3'-エクソヌクレアゼに対して高い耐性を示す.
- TPの改変は,RNAとのハイブリッド化時に軽度の不安定化を引き起こす.
- 光で標識されたTP-ODNは,効率的な細胞吸収を示しています.
- TP-ODNのサブセルラー分布は細胞タイプに依存し,HeLa細胞の核吸収が顕著である.
結論:
- トリアゾリルフォスフォナートODNは,オリゴヌクレオチド類同類物の汎用性のある新クラスを表しています.
- このクリック可能な化学アプローチは,新しい生物学的研究用試薬の設計に強力なツールを提供します.
- TP-改変されたODNは,さまざまなアプリケーションのために,安定性の向上と調節可能なセルラー吸収を提供します.
関連する概念動画
Phosphodiester Linkages
Overview
Phosphodiester bond forms when a phosphoric acid molecule (H3PO4) links with two hydroxyl groups (–OH) of two other molecules, forming two ester bonds. Two water molecules are released in this process. The phosphodiester bond is commonly found in nucleic acids (DNA and RNA) and plays a critical role in their structure and function.
Phosphodiester Bonds Link Nucleotides Together
DNA and RNA are polynucleotides or long chains of nucleotides that are linked together. A nucleotide is...
Phosphodiester bond forms when a phosphoric acid molecule (H3PO4) links with two hydroxyl groups (–OH) of two other molecules, forming two ester bonds. Two water molecules are released in this process. The phosphodiester bond is commonly found in nucleic acids (DNA and RNA) and plays a critical role in their structure and function.
Phosphodiester Bonds Link Nucleotides Together
DNA and RNA are polynucleotides or long chains of nucleotides that are linked together. A nucleotide is...
Nucleic Acid Structure
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...
DNA Structure
DNA has a double-helix structure. The...
The DNA Helix
Overview
The DNA Helix
Deoxyribonucleic acid, or DNA, is the genetic material responsible for passing traits from generation to generation in all organisms and most viruses. DNA is composed of two strands of nucleotides that wind around each other to form a spring-like structure called a double helix. However, the double helix is not perfectly symmetrical. Instead, there are regularly occurring grooves in the structure. The major groove occurs where the sugar-phosphate backbones are relatively far apart. This space...
Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Single-Strand DNA Binding Proteins
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...


