基酸DNA:一种多功能"点击"的骨干,用于基于DNA的生物应用
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
概括
研究人员开发了一种新的"点击"化学方法,以创建多用途的三基酸 (TP) DNA 类似物. 这些改性寡核酸 (ODN) 显示出更好的稳定性和细胞吸收,为生物研究提供了新的工具.
科学领域:
- 橄核酸的化学成分
- 化学生物学 化学生物学
- 生物技术是生物技术.
背景情况:
- 基于DNA的应用在细胞传递,非特异性影响,毒性和酶稳定性方面面临挑战.
- 现有的寡核酸类似物缺乏一种多功能方法来系统地解决这些问题.
研究的目的:
- 引入一种新的,可点击的化学物质,用于快速功能化DNA类似物.
- 合成和评估三基酸 (TP) 改性寡核酸 (ODN) 的生物特性.
主要方法:
- 开发一种与固相合成相容的基酸酸合成子.
- 使用"点击"化学来创建TP互核酸链接的后合成功能化.
- 混合骨干ODN与TP,酸盐或酸盐连接的自动固相合成,包括2'-OMe和LNA修改.
主要成果:
- TP链接对5'-和3'-外核酶具有很高的抗性.
- 在与RNA杂交时,TP修改会导致轻微的不稳定.
- 光标记的TP-ODN显示了有效的细胞吸收.
- TP-ODN的亚细胞分布取决于细胞类型,在HeLa细胞中显著的核吸收.
结论:
- 三基酸ODN代表了一类多功能新型的寡核酸相似物.
- 这种可点击化学方法为设计新生物研究试剂提供了强大的工具.
- 修改TP的ODN为各种应用提供了增强的稳定性和可调节的蜂接收.
相关概念视频
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.
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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...
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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...


