Novel Protecting Group Chemistry for Nucleic Acids

描述（由申请人提供）：将开发可在基因组方法中去除的耐热性核酸保护基团。沃森-克里克氢键中核碱基取代基的保护基团将被识别并用于阻止杂交。随后将耐热性组在高温下去除10-30分钟，作为基因组方法的一部分，将释放所需的核酸，并允许在受控的时间和温度条件下启动基因组方法。该技术有望消除由不匹配的杂化物、退火到非目标模板的引物、误引物和在室温下部分互补引物引起的非特异性反应和错误读数。主要目的是防止在样品制备过程中过早或错误地形成杂交核酸，从而导致序列识别的不良事件。具体目标是(1)发展可逆的、耐热的、共价的核酸修饰化学；(2)制备和表征适合于特定基因组方法的修饰核酸。核苷和小核苷酸将被用作模型化合物来研究候选化学物质和脱保护动力学。这些化学物质将应用于寡核苷酸引物、基因组DNA和三磷酸核苷进行评估。该项目将产生一类新的化合物和技术，用于健康相关的基因组方法，如SNP（单核苷酸多态性）分析和等位基因特异性寡核苷酸（ASO）杂交，这是应用于医学诊断和健康相关研究的DNA微阵列、PCR扩增和DNA测序的基础。商业应用包括修饰的核酸、特殊化学品和利用探针诊断、健康相关研究和病原体检测技术的新仪器。其他应用包括药物发现和药物反应基因研究（药物基因组学），生物防御和生物恐怖主义，可能需要“坚固”试剂，药物输送和治疗。

DESCRIPTION (provided by applicant): Thermolabile protecting groups of nucleic acids which can be removed during genomic methods will be developed. Protecting groups for nucleobase substituents involved in Watson-Crick hydrogen bonding will be identified and used to block hybridization. Subsequent removal of the thermolabile group at elevated temperature for 10-30 minutes, and as part of the genomic method, will release the desired nucleic acid and allow the genomic method to be initiated under controlled conditions of time and temperature. The technology is expected to eliminate non-specific reactions and false readings caused by mis-matched hybrids, primers annealed to non-target template, mis-priming, and partially complementary primers at ambient temperature. The primary objective is to prevent premature or erroneous formation of hybridized nucleic acids during sample preparation procedures that lead to undesirable events of sequence recognition. The specific aims are (1) development of chemistry for reversible, thermolabile, covalent modification of nucleic acids and (2) preparation and characterization of modified nucleic acids suitable for specific genomic methods. Nucleosides and small nucleotides will be used as model compounds to investigate candidate chemistries and the kinetics of deprotection. These chemistries will be applied to and evaluated with oligonucleotide primers, genomic DNA, and nucleoside triphosphates. This project will result in a new class of compounds and technology for health related applications in genomic methods such as SNP (single nucleotide polymorphism) analysis and allele-specific oligonucleotide (ASO) hybridization which are the basis of DNA microarrays, PCR amplification, and DNA sequencing applied to medical diagnostics and health related research. Commercial applications include modified nucleic acids, specialty chemicals, and new instrumentation for utilizing the technology in probe based diagnostics, health related research, and pathogen detection. Other applications include drug discovery and the study of drug response genes (pharmacogenomics), biodefense and bioterrorism where 'ruggedized' reagents may be needed, drug delivery, and therapeutics.