An Antisense Design and Simulation Platform

反义设计和仿真平台

• 批准号: 8058243
• 负责人: William J. Kennelly
• 金额: $ 13.99万
• 依托单位: DNA SOFTWARE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2011-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058243
• 关键词: Algorithms; Anti-Sense Probes; Antisense Oligonucleotides; Antisense Technology; Applications Grants; Binding; Biotin; Cardiovascular Diseases; Computer Simulation; Computer software; Databases; Dengue; Development; Diabetes Mellitus; Diagnostic; Engineering; Evaluation; Evaluation Research; Expenditure; Free Energy; Funding; Genomics; HIV; Hepatitis A; Hepatitis C; Human; Immunologic Deficiency Syndromes; Kinetics; Label; Laboratories; Libraries; Literature; Malignant Neoplasms; Measles; Measurement; Modeling; Molecular; Muscular Dystrophies; Oligonucleotides; Paramyxovirus; Phase; Public Health; RNA; Research; Research Personnel; Severe Acute Respiratory Syndrome; Small Business Innovation Research Grant; Software Tools; System; Temperature; Testing; Thermodynamics; Time; Update; Viral; Visual; West Nile virus; Work; analog; design; fluorophore; human disease; mathematical model; meetings; melting; phosphorothioate; research study; simulation; success; tool

DESCRIPTION (provided by applicant): In this grant proposal "An Automated Antisense Design and Simulation Platform", a comprehensive molecular diagnostics software tool that is specific to the design and simulation of antisense oligonucleotide analog probes will be developed. The application of antisense technologies to the study of human diseases has been proven in the literature for cancer, immune deficiency disorders, diabetes, muscular dystrophy and cardiovascular disease, hepatitis A and C, HIV, SARS-coronovirus, Ebola, Dengue Fever, paramyxoviruses (measles), and the West Nile virus. However, the progress of directed antisense research has been slow due to the absence of antisense oligonucleotide analog thermodynamic and kinetic databases, and because the currently used rule-of-thumb design strategies rarely ever initially produce effective probes. Many researchers are then forced to design a small library of probes against the same genomic target to increase their likelihood of success, which is very costly in both time and financing. The proposed Antisense Design and Simulation Platform will enable the researchers of human diseases by providing a tool for the directed research and evaluation of human and viral genomic targets so that rationally designed antisense oligonucleotide analogs may be used as a molecular diagnostics tool. The developmental strategies for producing the proposed platform are to update existing thermodynamic and kinetic databases and to deploy them on the Antisense Design and Simulation Platform, as outlined in the following specific aims: Aim 1.1: Perform 16 thermodynamic melts each for the fluorophore, biotin and quencher labeled PNA and morpholino antisense probes, and for the phosphorothioate/LNA antisense gap-mer probes to demonstrate the feasibility of applying the nearest-neighbor thermodynamic model to these systems. Aim 1.2: Add the thermodynamic parameters determined in Aim 1.1 to the existing PCR platform Visual OMP to demonstrate the feasibility of applying design and simulation algorithms to modified antisense oligonucleotide analogs. Aim 2.1: Complete the thermodynamic library for the fluorophore, and quencher labeled PNA and morpholino antisense probes, and for the phosphorothioate/LNA antisense gap-mer probes. Aim 2.2: Perform kinetics experiments on modified morpholino/RNA and PNA/RNA duplexes, and develop predictive mathematical models for the rates of Morpholino/RNA and PNA/RNA hybridization and unfolding. Aim 2.3: Engineer a fully automated Antisense Design and Simulation Platform that will allow researchers to design specific and sensitive antisense probes with minimal user inputs. By the end of this project a fully automated Antisense Design and Simulation Platform will have been designed, tested, and debugged, and made available to antisense researchers for the purpose of beta-testing the commercial product. PUBLIC HEALTH RELEVANCE: This project will directly impact public health by providing a comprehensive software tool for the design and simulation of antisense oligonucleotide analog probes, where no other comprehensive antisense software tools exist. The efficacy of applying antisense technologies to human diseases has been proven, using a trial-and-error approach to a number of human diseases, such as: cancer, immune deficiency disorders, diabetes, muscular dystrophy and cardiovascular disease, hepatitis A and C, HIV, SARS-coronovirus, Ebola, Dengue Fever, paramyxoviruses (measles), and the West Nile virus. The proposed Antisense Design and Simulation Platform will benefit the researchers of human diseases by providing an optimized molecular diagnostic tool for the directed research and rational evaluation of human and viral genomic targets.

描述（由申请人提供）：在本资助提案“自动化反义设计和模拟平台”中，将开发一种专门用于反义寡核苷酸类似物探针设计和模拟的综合分子诊断软件工具。反义技术在人类疾病研究中的应用已在癌症、免疫缺陷疾病、糖尿病、肌肉萎缩症和心血管疾病、甲型和丙型肝炎、HIV、SARS冠状病毒、埃博拉病毒、登革热、副粘病毒（麻疹）和西尼罗河病毒的文献中得到证实。然而，由于缺乏反义寡核苷酸类似物热力学和动力学数据库，并且由于目前使用的经验法则设计策略最初很少产生有效的探针，定向反义研究的进展缓慢。然后，许多研究人员被迫针对相同的基因组靶标设计一个小型的探针库，以增加他们成功的可能性，这在时间和资金上都非常昂贵。反义设计和模拟平台将为人类疾病的研究人员提供一个工具，用于人类和病毒基因组靶点的定向研究和评估，以便合理设计的反义寡核苷酸类似物可用作分子诊断工具。开发平台的发展战略是更新现有的热力学和动力学数据库，并将其部署在反义设计和模拟平台上，具体目标如下：目标1.1：对荧光团、生物素和猝灭剂标记的PNA和吗啉代反义探针进行16次热力学解链，以及硫代磷酸酯/LNA反义gap-mer探针，以证明将最近邻热力学模型应用于这些系统的可行性。目标1.2：将目标1.1中确定的热力学参数添加到现有PCR平台Visual OMP中，以证明将设计和模拟算法应用于修饰的反义寡核苷酸类似物的可行性。目标2.1：完成荧光团和淬灭剂标记的PNA和吗啉代反义探针以及硫代磷酸酯/LNA反义gap-mer探针的热力学文库。目标2.2：对修饰的吗啉/RNA和PNA/RNA双链体进行动力学实验，并开发吗啉/RNA和PNA/RNA杂交和解折叠速率的预测数学模型。目标2.3：设计一个完全自动化的反义设计和模拟平台，使研究人员能够以最少的用户输入设计特异性和敏感性的反义探针。到本项目结束时，一个全自动的反义设计和模拟平台将被设计，测试和调试，并提供给反义研究人员进行商业产品的β测试。 公共卫生关系：该项目将直接影响公众健康，提供一个全面的软件工具，设计和模拟反义寡核苷酸类似物探针，其中没有其他全面的反义软件工具存在。将反义技术应用于人类疾病的有效性已经被证明，使用试错法来治疗许多人类疾病，例如：癌症、免疫缺陷疾病、糖尿病、肌肉萎缩症和心血管疾病、甲型和丙型肝炎、艾滋病毒、SARS冠状病毒、埃博拉病毒、登革热、副粘病毒（麻疹）和西尼罗河病毒。该反义设计与模拟平台为人类和病毒基因组靶点的定向研究和合理评价提供了一个优化的分子诊断工具，将使人类疾病研究人员受益。