DNA-Scaffolded Peptides as High-Affinity Reagents

DNA 支架肽作为高亲和力试剂

• 批准号: 1506667
• 负责人: Jason Locklin
• 金额: $ 42万
• 依托单位: University of Georgia Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1506667&HistoricalAwards=false
• 关键词: DNA; Scaffolded; Peptides; Affinity; Reagents

Non-technical: This award by the Biomaterials program in the Division of Materials Research to University of Georgia is to develop a platform for antibody mimetics by scaffolding peptides on single stranded DNA. This award is cofunded by the Genetic Mechanisms program in the Division of Molecular & Cellular Biosciences in the Directorate for Biological Sciences. Antibodies are essential to facilitating discoveries in biomedical research; however, they have a variety of shortcomings that have elicited concerns over accessibility to high-quality antibodies for biomedical research. Alterative and more robust polymers for molecular recognition are needed to drive discovery efforts forward. Nucleic acid polymers have demonstrated immense promise as high-affinity reagents; however, their lack of chemical functionality limits their ability to compete with proteins as receptors. By merging the evolvable molecular scaffold of nucleic acid polymers with diverse protein functionality, the PI aims to evolve a new class of antibody mimetics for biomedical research, which may address the critical deficit of available antibodies for molecular targets. These research efforts will be coupled with educational and training outreach at the high school, undergraduate, and graduate levels in order to assist in the development of a future generation of scientists that are properly equipped to tackle interdisciplinary research problems.Technical: This proposal describes research to develop an in vitro selection platform for antibody mimetics by scaffolding peptides on DNA. Nucleic acid aptamers have provided numerous advantages over traditional antibodies; however, their functional group deficit limits their potential to match the performance of proteinogenic affinity reagents. By merging the evolvable molecular scaffold of nucleic acid polymers with protein functionality, this investigator will use this class of polymers to mimic the hetero- and multivalent approach nature uses at protein-protein interfaces. The investigator will use an enzymatic strategy to sequence-specifically incorporate unique peptide fragments along a DNA template. This approach will enable the concomitant evolution of the ssDNA scaffold and the identity of the displayed peptides to achieve molecular recognition of protein targets. If successful, this new class of biomimetic polymers could aid in addressing the deficit of high-affinity reagents for biomedical research. These research efforts will be coupled with educational and training outreach at the high school, undergraduate, and graduate levels.

非技术性：该奖项由格鲁吉亚大学材料研究部的生物材料项目授予，旨在通过在单链DNA上构建肽来开发抗体模拟物的平台。该奖项由生物科学理事会分子细胞生物科学部的遗传机制计划共同资助。抗体对于促进生物医学研究中的发现至关重要;然而，它们具有各种缺点，这些缺点引起了人们对生物医学研究中高质量抗体的可获得性的担忧。需要用于分子识别的可改变的和更稳健的聚合物来推动发现工作向前发展。核酸聚合物作为高亲和性试剂已经显示出巨大的前景;然而，它们缺乏化学功能性限制了它们作为受体与蛋白质竞争的能力。通过将核酸聚合物的可进化分子支架与不同的蛋白质功能相结合，PI旨在为生物医学研究开发一类新的抗体模拟物，这可能会解决分子靶标可用抗体的关键缺陷。这些研究工作将与高中、本科和研究生水平的教育和培训推广相结合，以帮助培养下一代科学家，他们有能力解决跨学科的研究问题。技术：本提案描述了通过在DNA上构建肽来开发抗体模拟物体外选择平台的研究。核酸适体提供了许多优于传统抗体的优点;然而，它们的官能团缺陷限制了它们与蛋白原亲和试剂的性能相匹配的潜力。通过将核酸聚合物的可进化分子支架与蛋白质功能性合并，本研究者将使用这类聚合物来模拟自然界在蛋白质-蛋白质界面上使用的异价和多价方法。研究者将使用酶促策略来将独特的肽片段沿着DNA模板进行序列特异性掺入。这种方法将使ssDNA支架的伴随进化和所展示的肽的同一性能够实现蛋白质靶标的分子识别。如果成功的话，这种新型仿生聚合物可以帮助解决生物医学研究中高亲和力试剂的不足。这些研究工作将与高中、本科和研究生阶段的教育和培训推广相结合。

项目成果

A High-Fidelity Codon Set for the T4 DNA Ligase-Catalyzed Polymerization of Modified Oligonucleotides

• DOI: 10.1021/acscombsci.5b00119
• 发表时间: 2015-12-01
• 期刊: ACS COMBINATORIAL SCIENCE
• 作者: Lei, Yi;Kong, Dehui;Hili, Ryan
• 通讯作者: Hili, Ryan

Sequence-Defined Scaffolding of Peptides on Nucleic Acid Polymers

• DOI: 10.1021/jacs.5b07675
• 发表时间: 2015-09-02
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Guo, Chun;Watkins, Christopher P.;Hili, Ryan
• 通讯作者: Hili, Ryan