EXTENDING THE SCOPE OF PNA WITH NON-STANDARD BASE PAIRS

使用非标准碱基对扩展 PNA 的范围

• 批准号: 6072136
• 负责人: GIDEON SHAPIRO
• 金额: $ 11.65万
• 依托单位: ERAGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6072136
• 关键词: biotechnology; high performance liquid chromatography; mass spectrometry; molecular biology information system; nucleic acid chemical synthesis; nucleic acids; nucleobase; peptide chemical synthesis; peptides; purines; pyrimidines

This research aims to expand the scope of Peptide Nucleic Acid (PNA) by adding artificial nucleobases which form stable non-standard base pairs to the currently available natural nucleobase PNA repertoire. PNAs based on an expanded nucleobase alphabet are anticipated to have significant potential in conjunction with the established properties of an expanded genetic information system (AEGIS). Incorporation of AEGIS non- standard DNA base pairs has improved bDNA diagnostic technology and led to a novel commercial DNA diagnostic product (bDNA 3.0 QuantiplexTM) with sensitivity comparable to PCR. Nevertheless, the scope of the DNA-AEGIS technology is limited by the difficult synthesis and attenuated chemical stability for several of the non-standard base reagents. PNA reagents and oligomers containing AEGIS non-standard nucleobases are anticipated to be more amenable to chemical synthesis and more stable. Furthermore, PNA has intrinisic properties which yield advantages for PNA:DNA based diagnostic systems relative to those based on DNA:DNA recognition (hybridization) complexes. These include greater stability and greater sensitivity to base mismatches for PNA:DNA duplexes relative to DNA:DNA hybrids. Taking the above together, inclusion of AEGIS non-standard nucleobases into PNA can be expected to greatly increase the scope and utility of both PNA and DNA-AEGIS technologies. Useful diagnostic applications can be already be conceived for the combination of these technologies. Specific examples include: coding for DNA microarrays, oligonucleotide ligation based DNA diagnostics. PROPOSED COMMERCIAL APPLICATIONS: Peptide Nucleic Acid (PNA) based on an expanded Watson:Crick rule based molecular recognition code will increase the scope and potential of PNA based diagnostic systems. Greater specificity for hybridization based assays taking advantage of DNA:PNA non-standard base pairing is expected. Specific applications with significant potential include the spatial address"zip coding" for DNA microarrays and other sandwich assays based on oligonucleotide ligation assay (OLA) in conjunction with the expanded code.

本研究旨在通过在现有的天然核碱基 PNA 库中添加形成稳定非标准碱基对的人工核碱基来扩展肽核酸 (PNA) 的范围。基于扩展核碱基字母表的 PNA 预计与扩展遗传信息系统 (AEGIS) 的既定特性相结合具有巨大潜力。 AEGIS 非标准 DNA 碱基对的结合改进了 bDNA 诊断技术，并产生了一种新型商业 DNA 诊断产品 (bDNA 3.0 QuantiplexTM)，其灵敏度可与 PCR 相媲美。然而，DNA-AEGIS 技术的范围受到合成困难和几种非标准碱试剂化学稳定性减弱的限制。含有 AEGIS 非标准核碱基的 PNA 试剂和寡聚物预计更适合化学合成并且更稳定。此外，PNA 具有固有的特性，相对于基于 DNA:DNA 识别（杂交）复合物的诊断系统，这些特性为基于 PNA:DNA 的诊断系统带来了优势。这些包括相对于 DNA:DNA 杂交体，PNA:DNA 双链体具有更高的稳定性和对碱基错配的更高敏感性。综上所述，将 AEGIS 非标准核碱基纳入 PNA 预计将大大增加 PNA 和 DNA-AEGIS 技术的范围和实用性。这些技术的组合已经可以构想出有用的诊断应用。具体例子包括：DNA 微阵列编码、基于寡核苷酸连接的 DNA 诊断。拟议的商业应用：基于扩展的 Watson:Crick 规则的分子识别代码的肽核酸 (PNA) 将扩大基于 PNA 的诊断系统的范围和潜力。 预计利用 DNA:PNA 非标准碱基配对的杂交测定具有更高的特异性。 具有巨大潜力的具体应用包括DNA微阵列的空间地址“zip编码”以及基于寡核苷酸连接测定(OLA)与扩展代码相结合的其他夹心测定。