Electronically Addressable DNA Assembly

电子可寻址 DNA 组装

• 批准号: 10697597
• 负责人: Matthew Taylor Holden
• 金额: $ 27.58万
• 依托单位: AVERY DIGITAL DATA, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10697597
• 关键词: Address; Binding; Biochemical Reaction; Buffers; Cells; Cellular biology; Complex; Custom; DNA; DNA Integration; DNA biosynthesis; DNA chemical synthesis; Data; Devices; Discipline; Electrodes; Electronics; Electrostatics; Enzymes; Evaluation; Excision; Frequencies; Generations; Genes; Genome; Geometry; Government; Human Genome; Immobilization; In Vitro; Infrastructure; Length; Measures; Microelectrodes; Modeling; Oligonucleotides; Persons; Phase; Primer Extension; Process; Production; Publishing; Reaction; Research; Running; Site; Solid; Specificity; Surface; System; Techniques; Technology; Translating; Work; analog; aspirate; cost; design; digital; electric field; electrical potential; fabrication; gene synthesis; genome-wide; improved; in vivo; interest; parallelization; prevent; self assembly; synthetic biology; voltage; whole genome

The ability to routinely synthesize entire genomes is one of the great remaining technical aspirations of the synthetic biology revolution. The complexity of designing the annealing reactions used by existing assembly workflows is a fundamental barrier to genome-scale DNA production. There is a practical limit to the number of distinct sequences which can reliably self-assemble into the desired construct in a single step, and traditional assembly reactions are typically limited in scale to tens or hundreds of oligonucleotides. Current parallel synthesis technologies can generate pools of over 1 million oligonucleotides, which is entirely inadequate for a genome-scale construction, yet far more complex than can be reliably assembled in a single reaction. Despite substantial interest in improving oligonucleotide synthesis throughput, such technologies cannot be leveraged fully without accompanying means of guiding their assembly. We have made advancements to generate oligonucleotides at a throughput 100× any published system. The proposed studies work to translate this advance (up to 100× enhancement) to the throughput of gene assembly. The underlying concept is to perform DNA synthesis on a substrate capable of controlling the localization of the oligonucleotides during their subsequent assembly. If successful, the approach would address a fundamental scalability mismatch between oligonucleotide synthesis and assembly, enabling a new generation of gene synthesis technologies capable of exceeding the scale of the human genome with a single synthesis run. This project may lead to a 100x parallelization increase and 100x cost decrease, and potentially increase assembly accuracy.

常规合成整个基因组的能力是人类最大的技术愿望之一， 合成生物学革命设计现有组件使用的退火反应的复杂性 工作流程是基因组规模DNA生产的根本障碍。有一个实际的限制， 可以在单个步骤中可靠地自组装成所需构建体的不同序列，以及传统的 组装反应的规模通常限于数十或数百个寡核苷酸。电流并联 合成技术可以产生超过100万个寡核苷酸的池，这对于一个生物体来说是完全不够的。 基因组规模的建设，但远比可以在一个单一的反应可靠地组装复杂。尽管 尽管对提高寡核苷酸合成通量有很大兴趣，但不能利用这些技术 完全没有伴随的引导其组装的装置。 我们已经取得了进展，以100倍于任何已发表系统的通量生成寡核苷酸。的 提出的研究工作将这一进展（高达100倍的增强）转化为基因组装的通量。 基本概念是在能够控制DNA的定位的基底上进行DNA合成。 寡核苷酸在其随后的组装过程中。如果成功的话，这种方法将解决一个根本的问题， 寡核苷酸合成和组装之间的可扩展性不匹配，使得新一代基因 一次合成就能超过人类基因组规模的合成技术。这 一个项目可能导致并行化增加100倍，成本降低100倍，并可能增加组装 精度