Electronically Addressable DNA Assembly

电子可寻址 DNA 组装

基本信息

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
项目类别：
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

项目摘要

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尚存的伟大技术愿望之一合成生物学革命。设计现有组装所使用的退火反应的复杂性。工作流程是基因组规模 DNA 生产的基本障碍，但实际数量存在限制。不同的序列可以在一个步骤中可靠地自组装成所需的构建体，而传统的当前并行的组装反应的规模通常限于数十或数百个寡核苷酸。合成技术可以生成超过 100 万个寡核苷酸库，这对于基因组规模的构建，但比在单个反应中可靠组装要复杂得多。对提高寡核苷酸合成通量有很大兴趣，因此无法利用此类技术完全没有附带的引导其组装的装置。我们已经取得了进展，能够以任何已发布系统 100 倍的吞吐量生成寡核苷酸。拟议的研究致力于将这一进步（高达 100 倍的增强）转化为基因组装的吞吐量。基本概念是在能够控制 DNA 定位的基质上进行 DNA 合成。如果成功，该方法将解决一个基本问题。寡核苷酸合成和组装之间的可扩展性不匹配，使新一代基因成为可能能够通过一次合成运行超越人类基因组规模的合成技术。该项目可能会导致并行度提高 100 倍，成本降低 100 倍，并可能增加装配量准确性。