在非核友性去保护条件下进行的寡二氧核酸合成
Shiyue Fang1, Komal Chillar1, Yipeng Yin1
1Department of Chemistry and Health Research Institute, Michigan Technological University, Houghton, Michigan.
Current protocols
|February 8, 2024
概括
一种新方法可以通过使用新型的保护组和裂变条件来合成敏感的寡氧核酸 (ODN). 这种方法可以为各种研究应用创建以前无法访问的DNA修改.
科学领域:
- 化学生物学 化学生物学
- 有机合成 有机合成
- 分子生物学分子生物学
背景情况:
- 标准的寡氧核酸 (ODN) 合成方法采用了与敏感功能组不相容的苛刻的基本和核性条件.
- 像N4-乙脱氧化素 (4acC),化等修饰在ODN合成过程中容易降解.
- 开发温和合成策略对于访问各种应用程序的修改ODN至关重要.
研究的目的:
- 描述一种用于合成敏感的寡度氧核酸 (ODN) 的新方案.
- 引入一种保护和链接策略,规避严格的剥夺保护条件.
- 为了使更广泛的修改ODN用于研究的合成.
主要方法:
- 使用一个1,3-dithian-2-yl-methoxycarbonyl (Dmoc) 保护组,特别是甲基替代的meDmoc变体,用于核基外循环胺.
- 通过使用Dmoc链接器将不断增长的ODN固定在坚实的支持上.
- 开发了一种使用近乎中性,非核性氧化条件的去保护和裂解策略.
- 引入了PEGylated Dmoc (pDmoc) 胺,以提高溶解度,并使更长的ODN的合成成为可能.
主要成果:
- 成功合成了包含核敏感功能组的ODN.
- 证明meDmoc和pDmoc策略允许在温和,非核爱的条件下进行ODN脱保护和裂变.
- 该pDmoc修改提高了固体支绑定ODN的可溶性,促进了较长序列的合成.
结论:
- 描述的meDmoc方法提供了一个强大的方法来合成以前难以访问的敏感ODN.
- 该协议扩展了用于表观遗传学,药物开发和纳米技术创建改性核酸的工具包.
- 该方法有望促进在需要定制设计的ODN的不同领域的研究.
相关概念视频
Proofreading
6.3K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
Errors During Replication are Corrected by the DNA Polymerase...
6.3K
Translesion DNA Polymerases
10.0K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
10.0K
ATP and Macromolecule Synthesis
5.6K
Biological macromolecules are organic compounds, predominantly composed of carbon atoms. The carbon atoms are covalently bonded with hydrogen, oxygen, nitrogen, and other minor elements. There are four major biological macromolecule classes: carbohydrates, lipids, proteins, and nucleic acids.
Most macromolecules are composed of single subunits, or building blocks, called monomers. The monomers combine with each other using covalent bonds to form larger molecules known as polymers.
Conversion of...
Most macromolecules are composed of single subunits, or building blocks, called monomers. The monomers combine with each other using covalent bonds to form larger molecules known as polymers.
Conversion of...
5.6K
Lagging Strand Synthesis
51.6K
During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
51.6K
Homologous Recombination
50.5K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.5K
Overview of DNA Repair
31.0K
In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
Chemically...
Chemically...
31.0K


