High Accuracy Nanopore Sequencing.

高精度纳米孔测序。

基本信息

  • 批准号:
    10457928
  • 负责人:
  • 金额:
    $ 88.16万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2009
  • 资助国家:
    美国
  • 起止时间:
    2009-09-23 至 2025-05-31
  • 项目状态:
    未结题

项目摘要

PROJECT SUMMARY The objective of this proposal is to unlock the full potential of nanopore sequencing. This sequencing method has transformative intrinsic qualities, such as long read length, direct epigenetic modification detection, fast sample-to-answer, portability and low cost- to-entry, but relatively low sequencing accuracy remains a significant drawback. With NHGRI funding, we have built a team that has played a pivotal role in developing nanopore sequencing, first engineering the highly sensitive pore MspA to provide single-nucleotide resolution and then combining it with enzyme control for the first proof-of-concept of nanopore sequencing. Recently, we developed a hybrid- voltage-enzyme control method that provided a substantial increase in nanopore sequencing accuracy. Concurrently, we developed a nanopore single-molecule tool that measures the kinetics of enzymes which move along DNA or RNA at unprecedented detail. Here we build on our previous research and propose well-founded and innovative methods to further increase the accuracy of nanopore sequencing with the ultimate goal of a single-passage base calling accuracy of around 99%. Our specific aims are (1) to use sequence-dependent enzyme kinetics to create a second reader in the nanopore system that runs in tandem to the ion current reader. This reader will provide sequence information that is independent of the ion current reader. We will engineer enzymes with accentuated sequence-dependent kinetics. (2) We will systematically map the sequence-dependent enzyme kinetics and splice this information into novel base calling algorithms. In addition, we will measure the ion current of all four different orientations of the pore-enzyme-DNA complex to maximize the accuracy with this new method. (3) We will engineer better sequence specificity into the ion current reader by creating a robust platform that permits asymmetric assembly of MspA. We will then design MspA with an asymmetric pore constriction to reduce Brownian motion of the DNA and enhance nucleotide recognition. Our team's success to date has enabled us to form partnerships with prominent collaborators and to gain support from many excellent labs in academia and industry, whose expertise assists us in each step forward. We will work with our partner labs to complete the aims outlined in this proposal.
项目总结

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(15)

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JENS GUNDLACH其他文献

JENS GUNDLACH的其他文献

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{{ truncateString('JENS GUNDLACH', 18)}}的其他基金

Nanopore sequencing of DNA with MspA
使用 MspA 进行 DNA 纳米孔测序
  • 批准号:
    8728979
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
High Accuracy Nanopore Sequencing.
高精度纳米孔测序。
  • 批准号:
    10214247
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
High Accuracy Nanopore Sequencing
高精度纳米孔测序
  • 批准号:
    9980937
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
High Accuracy Nanopore Sequencing
高精度纳米孔测序
  • 批准号:
    10267319
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
Nanopore sequencing of DNA with MspA
使用 MspA 进行 DNA 纳米孔测序
  • 批准号:
    8572105
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
Nanopore sequencing of DNA with MspA
使用 MspA 进行 DNA 纳米孔测序
  • 批准号:
    8903338
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
Nanopore sequencing of DNA with MspA
使用 MspA 进行 DNA 纳米孔测序
  • 批准号:
    8910775
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
Nanopore sequencing of DNA with MspA
使用 MspA 进行 DNA 纳米孔测序
  • 批准号:
    9145099
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
High Accuracy Nanopore Sequencing.
高精度纳米孔测序。
  • 批准号:
    10631962
  • 财政年份:
    2009
  • 资助金额:
    $ 88.16万
  • 项目类别:
Engineering MspA for Nanopore Sequencing
用于纳米孔测序的工程 MspA
  • 批准号:
    7192749
  • 财政年份:
    2006
  • 资助金额:
    $ 88.16万
  • 项目类别:

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