Massively parallel linearization and chemical treatment of DNA molecules using tunable nanoscale confinement

基因分析，如DNA测序，已成为健康科学研究以及疾病预后和诊断的既定技术。然而，现代测序方法面临着相当大的挑战，带来了关键技术的发展机会。例如，BCC Research预计到2018年全球测序市场将增长至近117亿美元，2013年至2018年的年增长率为21.2%。作为当今测序市场的主要股东，Illumina的技术依赖于将DNA分解成数百万个小片段，比基因组小许多数量级。这种断裂过程通常导致昂贵且容易出错的基因组重组过程，以及长范围基因组信息的丢失，例如癌症中发生的单倍型相位、拷贝数变异和结构重排。以Pacific Biosciences为例，竞争性的长分子技术受到高错误率的困扰，这使得它们的结果在计算上不切实际，无法在利基应用之外使用。该提案的目的是研究、开发和合成新技术，以比现有技术更长的DNA链测序，并降低错误率。我们结合了联合收割机在纳米级物理和单分子显微镜，微机电系统（MEMS）工程和标记化学的关键专业知识。这一提议启动了三个世界专家之间的关系：麦吉尔大学的莱斯利实验室开创了新颖的单分子操纵和可视化"CLiC"技术，该技术对轻轻解开，线性化和分析长DNA链至关重要; Micralyne是加拿大领先的微流体和MEMS铸造厂，拥有广泛的制造专业知识，这是实现莱斯利技术的关键; Genetics已经开发出化学处理方法来标记DNA碱基对，以便灵敏地读出。该项目提供的资源、培训、见解和技术将为加拿大生物技术部门的发展培养高素质的人才，创造就业机会和收入，例如，与该技术相关的消耗品的制造将在Micralyne（加拿大埃德蒙顿）进行。

Genetic analysis, such as DNA sequencing, has become an established technology for health science research, as well as disease prognosis and diagnosis. However, modern sequencing methods face considerable challenges, presenting key technology development opportunities. For instance, BCC Research expects the global sequencing market to grow to nearly $11.7 billion by 2018 and register an annual growth rate of 21.2% from 2013 to 2018. The technology underlying today's sequencing market dominant shareholder, Illumina, relies on breaking DNA into millions of tiny pieces, many orders of magnitude smaller than the genome. This fracturing process typically leads to a costly and error-prone process of reassembling the genome, and loss of long-range genomic information such as haplotype phase, copy number variation, and structural rearrangements that occur in cancer. Competing long-molecule technologies, as exemplified by Pacific Biosciences, are plagued by high error rates that render their results computationally impractical to use beyond niche applications. The aim of this proposal is to research, develop, and synthesize new technologies to sequence longer DNA strands than current technology and with lower error rates. We combine key expertise in nanoscale physics and single-molecule microscopy, microelectromechanical systems (MEMS) engineering, and labeling chemistry. This proposal kickstarts a relationship between three world-experts: the Leslie Laboratory at McGill University has pioneered novel single-molecule manipulation and visualization "CLiC" technology critical to gently untangling, linearizing and analyzing long DNA strands; Micralyne is a leading microfluidics and MEMS foundry in Canada, with extensive fabrication expertise key to miniaturizing Leslie's techniques; ZSGenetics has developed chemical processing approaches to label DNA base pairs for sensitive readout. The resources, training, insights and technologies delivered by this project would train high-quality personnel for growing the biotechnology sector in Canada, generating employment and revenue e.g. fabrication of consumables associated with the technology would be anchored in Micralyne (Edmonton, Canada).