CAREER: Probing Stem Cell Differentiation With Synthetic Biology

PI：Deans，塔拉 提案编号：1554017干细胞的需求预计将继续上升，由于其预期的能力，以治疗疾病，用于新的诊断技术，并用于药物筛选。然而，在实现这些目标之前，需要更好地了解调节其命运的机制。干细胞整合周围的许多信号，并根据这些输入执行细胞行为。这些属性可以使用合成生物学来利用和操纵，以动态模式严格控制基因表达，此外还可以编程细胞来感知和记录其微环境的变化。 一旦成功完成，这项工作将使参与干细胞命运的决定，最终可能被利用到直接分化的机制有更好的理解。内在（转录因子表达），以及外在（环境）机制被认为是参与调节干细胞自我更新，并致力于分化成更专门的细胞类型。这种内在和外在的线索之间的相互作用，在分化提出了挑战，研究其增殖和终末分化的机制。这些挑战可以通过基因电路来解决。这项研究的完成将产生工具来查询不同的时间，身份和细胞命运的内部和外部信号的相互作用。这将通过开发使用遗传电路探测干细胞分化的策略来实现，其目标如下：（1）扩展用于动态控制哺乳动物细胞中的基因表达的遗传工具箱，（2）通过将遗传电路连接到已知参与细胞命运选择的重要受体来评估干细胞分化时细胞外微环境的变化，以及（3）使用这些工具来探测干细胞分化。 这项研究的所有结果都将提供给研究界。这项研究计划的另一个重要的广泛影响领域是教育和STEM领域的推广，包括小学，高中和本科生。

PI: Deans, Tara Proposal Number: 1554017The demand for stem cells is anticipated to continue to rise due to their expected ability to treat disease, use in novel diagnostic technologies, and for pharmaceutical screening. However, before these goals can be realized, a better understanding of the mechanisms regulating their fate is required. Stem cells integrate the many signals that surround them and execute cellular behaviors based on these inputs. These attributes can be harnessed and manipulated using synthetic biology to tightly control gene expression in dynamic patterns, in addition to programming cells to sense and record changes in their microenvironment. Upon successful completion, this work will enable a better understanding of the mechanisms involved in stem cell fate decisions that may eventually be exploited to direct differentiation.Both intrinsic (transcription factor expression), as well as extrinsic (environmental) mechanisms are thought to be involved in the regulation of stem cell self-renewal, and their commitment to differentiate into more specialized cell types. This interplay between intrinsic and extrinsic cues in differentiation poses challenges to studying the mechanisms involved in their proliferation and terminal differentiation. These challenges can be addressed using genetic circuits. The completion of this research will produce tools to query the different timings, identities, and interactions of the internal and external signals of cell fate. This will be accomplished by developing strategies for probing stem cell differentiation using genetic circuits with the following objectives: (1) to expand the genetic toolbox for dynamically controlling gene expression in mammalian cells, (2) to assess changes in the extracellular microenvironment as stem cells differentiate by connecting genetic circuits to important receptors known to be involved in cell fate choices, and (3) to use these tools to probe stem cell differentiation. All of the results of this research will be made accessible to the research community. An additional important broad impact area of this research program is education and outreach of STEM fields at a variety of levels including elementary school, high school, and undergraduate students.