Multipurpose reversible terminators

多用途可逆终结器

• 批准号: 8529712
• 负责人: Daniel Hutter
• 金额: $ 26.2万
• 依托单位: FIREBIRD BIOMOLECULAR SCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-17 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8529712
• 关键词: Abbreviations; American; Architecture; Base Sequence; Biological; Biological Assay; Biology; Buffers; C-glycoside; Cicatrix; Cleaved cell; Complex; Contracts; DNA; DNA Sequence; DNA analysis; DNA-Directed DNA Polymerase; Data; Deoxyribonucleotides; Detection; Development; Diagnostic; Disulfides; Emulsions; Enzymes; Excision; Family; Foundations; Funding; Genetic; Genomics; Glycols; Grant; Hereditary Disease; Human; Information Systems; Left; Length; Licensing; Link; Market Research; Medical; Medicine; Modeling; Modification; Molecular Evolution; Mutation; Natural regeneration; Nitrogen; Nitrous Acid; Nucleic Acids; Nucleic acid sequencing; Occupations; Oils; Oligonucleotides; Phase; Polymerase; Polymorphism Analysis; Primer Extension; Production; Pyrimidine; RNA-Directed DNA Polymerase; Reading; Reagent; Recovery; Research; Risk; Sales; Sampling; Science; Scientist; Series; Side; Single Nucleotide Polymorphism; Small Business Technology Transfer Research; Sodium Nitrite; Structure; Sulfhydryl Compounds; Synthesis Chemistry; System; Technology Transfer; Therapeutic; Time; Tissues; Transferase; Water; Work; appendage; base; dideoxynucleotide; effectiveness research; functional group; innovation; metaperiodate; molecular recognition; mutant; next generation; nucleic acid analog; nucleoside triphosphate; phosphoramidite; public health relevance; software development; tool; tripolyphosphate; two-dimensional

DESCRIPTION (provided by applicant): This Phase 2 STTR application will allow the Foundation for Applied Molecular Evolution (FfAME) to complete its transfer of technology to Firebird Biomolecular Sciences LLC, allowing Firebird to commercialize nucleoside triphosphates where the 3'-OH groups are blocked by a 3'-ONH2 group. Triphosphates having this modification are accepted by DNA polymerases and other enzymes. After being incorporated at the 3'-end of a growing oligonucleotide, the 3'-ONH2 group blocks extension (just as a 2',3'-dideoxynucleotide does). However, this blockage is reversible; buffered sodium nitrite rapidly cleaves the O-NH2 bond to generate the natural 3'-OH group, yielding under mild conditions a primer that can again be extended by DNA polymerase. Further, the 3'-ONH2 group offers a handle that allows the terminated DNA to selectively react with certain functional groups, allowing for the recovery of the extended, terminated DNA from complex biological mixtures. This is useful in many architectures that analyze DNA, including "next generation" sequencing that promises to personalize diagnostics and therapeutics, tools to detect single nucleotide polymorphisms, strategies to do "effectiveness research" for medical treatments, and other architectures that have increasing complexity and risk. Therefore, the products to be developed here have multiple applications in a variety of research settings, genetic disease discovery, and human diagnostics. This Phase 2 project, together with Firebird's internal funding, will build a production pipeline to prepare 12 different triphosphates carrying the reversible terminator, as well as four phosphoramidites to support kits that use these. The synthetic chemistry developed in the Phase 1 period will support this pipeline. Further, tools used in the Phase I period to develop polymerases to handle unnatural nucleoside triphosphates will be applied to create still better polymerases for assays that exploit our triphosphates. Finally, kits for DNA analysis will be produced that exploit these reversible terminators, in many cases with other reagent innovations that are proprietary to FfAME, Firebird, and others. Several customers are already exploiting these compounds. These include foreign customers, whose purchase of these reagents is creating American jobs through the sale of products internationally. Growing its portfolio of nucleic acid analogs underlies the next phase in the development of Firebird as a corporation. PUBLIC HEALTH RELEVANCE: Firebird Biomolecular Sciences LLC commercializes innovative reagents, polymerases that exploit them, and assays and architectures based on them, for the detection, analysis and sequencing of nucleic acids. Firebird also develops software to exploit genomic sequence data, and what it means in biology and medicine. This Phase 2 STTR project will create over two dozen jobs through the development of a series of reversible terminators suitable for next generation DNA sequencing, SNP detection, and DNA capture.

描述（由申请人提供）：此阶段2 STTR应用将允许应用分子进化（FFAME）的基础完成其技术转移到Firebird Biomolocular Sciences LLC，从而使Firebird可以将核苷三磷酸盐商业化，其中3'-OH基团被3'-on-ONH2组阻止。 DNA聚合酶和其他酶接受了具有这种修饰的三磷酸盐。在生长寡核苷酸的3'末端掺入后，3'-ONH2组阻止了延伸（就像2'，3'-二氧基核苷酸一样）。但是，这种阻塞是可逆的。缓冲钠硝酸钠迅速裂解O-NH2键以产生天然3'-OH基团，在轻度条件下产生底漆，可以再次通过DNA聚合酶扩展。此外，3'-ONH2组提供了一个手柄，允许终止的DNA与某些官能团有选择性反应，从而可以从复杂的生物混合物中回收扩展的，终止的DNA。这在分析DNA的许多架构中很有用，包括有望个性化诊断和治疗学的“下一代”测序，检测单核苷酸多态性的工具，对医疗治疗进行“有效研究”的策略以及其他具有复杂性和风险增加的建筑。因此，这里要开发的产品在各种研究环境，遗传疾病发现和人类诊断中都有多个应用。该第2阶段的项目以及Firebird的内部资金将建立一条生产管道，以准备12种带有可逆终结者的三磷酸盐，以及四个磷光素，以支持使用这些套件的工具包。在第1期时期开发的合成化学将支持该管道。此外，将在I期时期用于开发聚合酶来处理非天然核苷三磷酸盐的工具，以创建更好的聚合酶，以利用我们的三磷酸化的测定法。最后，将生产用于DNA分析的套件，以利用这些可逆终止者，在许多情况下，具有其他试剂创新，这些创新具有ffame，Firebird等。一些客户已经在利用这些化合物。其中包括外国客户，他们购买这些试剂是通过国际出售产品来创造美国就业机会。将其核酸类似物的投资组合发展为Firebird作为公司发展的下一阶段。 公共卫生相关性：Firebird Biolocular Sciences LLC将创新试剂，利用它们的聚合酶以及基于它们的测定和结构进行商业化，以进行核酸的检测，分析和测序。 Firebird还开发了用于利用基因组序列数据及其在生物学和医学中的意义的软件。该第2阶段的STTR项目将通过开发一系列适合下一代DNA测序，SNP检测和DNA捕获的可逆终结器来创造超过两打作业。