CAREER: Riboswitch-based whole-cell biosensors to detect small organic molecules: A combined educational and research plan

职业：基于核糖开关的全细胞生物传感器检测有机小分子：教育和研究相结合的计划

• 批准号: 1258307
• 负责人: Jane Liu
• 金额: $ 40.48万
• 依托单位: Pomona College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1258307&HistoricalAwards=false
• 关键词: CAREER; Riboswitch; based; whole; cell

1148202LiuDrew UniversityAlthough small organic molecules are essential to all biological systems, there remains a need for tools that allow scientists to measure the concentration of any given small molecule in a biologically relevant setting. The project leader's long-term research goal is to develop a general method to rapidly and easily monitor small molecules relevant to the study of biology and the environment. The objective of this proposal is to engineer riboswitch-based biosensors to monitor intracellular concentrations of small organic molecules. Riboswitches are naturally occurring non-coding RNA elements that act as direct sensors of diverse small molecules and can signal molecular recognition through altered protein expression. The rationale for this project is that the demonstrated plasticity of riboswitches coupled with the utility of genetic selections can form the basis of a powerful method for engineering whole-cell biosensors for small molecules. The proposed work sets out to demonstrate the utility of riboswitch-based biosensors by using a naturally occurring riboswitch to identify environmental factors that may contribute to the metabolism of an important signaling molecule, cyclic diguanylate, in Vibrio cholerae. To demonstrate how genetic selections can modify and improve natural riboswitches for use in biosensors, the V. cholerae-specific cyclic diguanylate riboswitch will also be re- engineered to generate a new riboswitch that can monitor cyclic diguanylate in any bacterial species. The potential of such a tool is immense in terms of its ability to advance the entire field of microbiology research. Finally, to demonstrate the full range and potential of this approach, a riboswitch-based biosensor for polychlorinated biphenyls will be engineered through directed evolution. The successful completion of this aim would produce a biosensor that can provide a sensitive and inexpensive means to monitor bioavailable and bioaccessible levels of a persistent organic pollutant. These models will define the precision with which riboswitch-based biosensors can be generated and will speak to their broad utility. This contribution is transformative because it is expected to provide a system that will be able to rapidly detect and report the intracellular level of virtually any cell permeant small molecule of interest. The engineering of biosensors and the use of non-coding RNAs as research tools, moreover, present an exciting frontier for a diverse group of budding scientists. The project leader is committed to graduating scientifically literate undergraduates and motivating students to pursue life-long careers in science; this project directly addresses these career goals. Over a dozen undergraduate summer research positions will be funded over the five-year duration of the project. Students working on this project will communicate their results to the scientific community as co-authors of journal articles and as presenters at professional meetings. Student researchers will also be asked to contribute to a blog which seeks to communicate to other undergraduate and high school students what science research is and what it produces. As part of this project, authentic research experiences will be introduced into the curriculum for both majors and non-majors. In addition to providing experiential learning on-campus, the project leader is also initiating a partnership with a research-intensive institution to create off-campus research opportunities for her undergraduate students.

1148202 liudrew Universityal，尽管小有机分子对于所有生物系统都是必不可少的，但仍需要工具，使科学家可以在生物学相关的环境中测量任何给定的小分子的浓度。项目负责人的长期研究目标是开发一种通用方法，以快速，轻松地监测与生物学和环境研究相关的小分子。该建议的目的是设计基于核糖开关的生物传感器来监测细胞内小有机分子的浓度。核糖开关是自然存在的非编码RNA元件，它们充当不同小分子的直接传感器，可以通过改变蛋白表达来信号分子识别。该项目的基本原理是，结合遗传选择的核糖开关的可塑性可以构成一种强大的方法来设计小分子的全细胞生物传感器。拟议的工作旨在通过使用天然存在的核糖开关来识别可能有助于重要信号分子的代谢，环状二甘氨酸，在Vibrio Cholerae中的代谢，以证明基于核糖开关的生物传感器的实用性。为了证明遗传选择如何修改和改善用于生物传感器的天然核糖开关，还将重新设计霍乱特异性环状二甘氨酸核糖开关，以产生一种新的核糖开关，该新核糖开关可以监测任何细菌种类中的环状二烷基化。这种工具的潜力在推进整个微生物研究领域的能力上是巨大的。最后，为了证明这种方法的全部范围和潜力，将通过定向进化来设计基于核糖开关的多氯联苯生物传感器。该目标的成功完成将产生一种生物传感器，该生物传感器可以提供一种敏感且廉价的手段，以监测持续性有机污染物的可生物利用和可生物接收水平。这些模型将定义可以生成基于核糖开关的生物传感器的精度，并将与其广泛的实用程序有关。这种贡献具有变革性，因为它有望提供一个能够快速检测和报告几乎任何细胞渗透性小分子的细胞内水平的系统。此外，生物传感器的工程和使用非编码RNA作为研究工具，为各种各样的崭露头角的科学家提供了令人兴奋的边界。项目负责人致力于毕业科学识字的大学生，并激励学生从事科学终身职业。该项目直接解决了这些职业目标。 在该项目的五年期间，将资助十多个本科生的夏季研究职位。从事该项目的学生将作为期刊文章的合着者和专业会议的主持人传达其结果。还将要求学生研究人员为一个博客做出贡献，该博客旨在与其他本科和高中生交流什么是科学研究以及它的产生。作为该项目的一部分，将向专业和非律师提供真实的研究经验。除了提供体验式学习校园外，项目负责人还与研究密集型机构建立了合作伙伴关系，为她的本科生创造了校外研究机会。