Automated long DNA synthesis technology

描述（由申请人提供）：准确和高通量合成长DNA序列的能力是最终理解复杂基因组组织和功能问题的关键。有了这样的能力，就可以通过系统地测试设计和合成的基因或染色体序列的生物学效应来研究任何潜在基因和染色体调控元件的身份、位置和功能，其中许多是远程的。从头合成，而不是对天然基因组元素的简单重排，给了研究人员完全的自由来测试设计的新序列或精细序列变异对基因组功能的影响。然而，现有的基因合成技术仍然依赖于昂贵和繁琐的大规模DNA寡核苷酸合成和人工基因组装程序。本课题旨在开发集成的、基于微流控和微阵列的从头基因合成技术平台来填补这一空白。这个自动化的基因合成平台能够准确地将设计的基因序列“写入”双链DNA分子，为克隆、表达和其他下游应用做好准备。与热循环仪一样，自动化基因合成器也将得到广泛应用，成为生命科学研究人员不可或缺的工具。除了基因组结构和功能研究之外，该技术还将使许多目前难以或效率极低的生物医学研究成为可能，例如大规模合成基因组学，遗传电路和代谢途径的构建，新蛋白质，酶，抗体和其他药物的设计，进化和优化。

DESCRIPTION (provided by applicant): The capability of accurate and high-throughput synthesis of long DNA sequences holds the key to ultimate understanding of complex genome organization and function issues. With such capabilities, the identity, position and function of any potential gene and chromosomal regulation elements, many of which are long-range, can be studied by systematically testing the biological effects of designed and synthesized gene or chromosomal sequences. De novo synthesis, instead of simple rearrangements of natural genomic elements, gives researchers total freedom to test the effects of designed novel sequences or fine sequence variations on genome function. However, existing gene synthesis technology still relies on costly and cumbersome macro- scale DNA oligonucleotide synthesis and manual gene assembly procedures. This proposal aims to develop integrated, microfluidics and microarray-based de novo gene synthesis technology platform to fill this gap. This automated gene synthesis platform is able to accurately "write" designed gene sequences into double-stranded DNA molecules, ready for cloning, expression and other down-stream applications. Like thermal cyclers, automated gene synthesizers will be widely used and become an indispensable tool for life science researchers. In addition to genome structure and function studies, the technology will enable many types of biomedical research which are currently difficult or extremely inefficient to perform, such as large scale synthetic genomics, construction of genetic circuits and metabolic pathways, design, evolution and optimization of new proteins, enzymes, antibodies and other pharmaceuticals. PUBLIC HEALTH RELEVANCE: This proposal aims to develop accurate automated and low cost de novo gene synthesis technology platform. Such a technology would be extremely useful for studying complex genome organization and function issues. In addition, the technology will enable many types of biomedical research which are currently difficult or extremely inefficient to perform, and will lead to better understanding of human biology and diseases.