A low-cost benchtop oligonucleotide synthesizer using inkjet enzymatic DNA synthesis

使用喷墨酶法 DNA 合成的低成本台式寡核苷酸合成仪

基本信息

批准号：
10213239
负责人：
ADRIAN HORGAN
金额：
$ 73.45万
依托单位：
DNA SCRIPT
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-06-30
项目状态：
已结题

项目摘要

ABSTRACT The objective of this proposal is to develop a benchtop, inkjet-based, oligonucleotide synthesizer using aqueous, enzymatic DNA synthesis technologies for the parallel synthesis of up to 1 million high-quality oligonucleotides per day. The system will deliver >10-fold improvements in the key metrics of cost, quality, and throughput compared to existing technologies. The synthesis biochemistry will utilize engineered terminal deoxynucleotidyl transferase enzymes that have been optimized for incorporation of 3’ aminoxy reversible terminator nucleotides. This approach has several advantages, including proven performance in the synthesis of oligonucleotides up to 300 nucleotides on microbeads. Here, we will adapt the technology to enable DNA synthesis on functionalized microscope slides using inkjets for precise reagent delivery. A multidisciplinary approach will be followed to advance toward the goals in terms of engineering for instrumentation development, biochemistry and protocol optimization, and performance benchmarking of the synthesized oligonucleotides. During the first year, we will build an initial testbed to enable verification and optimization of individual components, materials, reagents, and protocols. In the second year, we will build a fully automated prototype instrument, and refine the protocols and software to allow synthesis of long oligonucleotides. Finally, Year-3, we will build multiple benchtop instruments and work with partner laboratories to test the oligonucleotides in several ‘real world’ applications. Subsequent commercialization of the instruments will democratize access to rapid high-density oligonucleotide synthesis, and revolutionize approaches to a wide variety of synthetic biology and functional genomics applications.

抽象的该建议的目的是开发台式，基于喷墨的寡核苷酸合成器使用水性，酶促DNA合成技术，以平行合成多达100万每天高质量的寡核苷酸。该系统将在密钥中提供> 10倍的改进与现有技术相比，成本，质量和吞吐量的指标。综合生物化学将利用具有工程的末端脱氧核苷酸转移酶已针对3'aminooxy可逆终结核苷酸的保险进行了优化。这种方法具有几个优势，包括在合成寡核苷酸的合成中的经验证明微粒上的300个核苷酸。在这里，我们将适应该技术以启用DNA合成使用喷墨以进行精确试剂递送的功能化显微镜载玻片。多学科将遵循方法，以迈向仪器工程学的目标开发，生物化学和协议优化以及性能基准合成的寡核苷酸。在第一年，我们将建立一个初始测试以启用验证和优化各个组件，材料，试剂和协议。在第二年，我们将建立一个完全自动化的原型工具，并完善协议和允许合成长寡核苷酸的软件。最后，3年级，我们将建造多个台式仪器，并与合作伙伴实验室合作测试几种寡核苷酸 “现实世界”应用。随后的工具商业化将民主化获取快速高密度寡核苷酸的合成，并彻底改变了范围的方法多种合成生物学和功能基因组学应用。