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Mammalian synthetic biology: From discovery to therapy.

机译：哺乳动物合成生物学：从发现到治疗。

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摘要

The nascent field of mammalian synthetic biology holds the promise to revolutionize personalized medicine. The use of engineered genetic circuits for genome-wide screens, detecting and reporting molecular signatures, and targeting genetic mutations can redefine personalized medicine with specific applications ranging from discovery to therapy.;Genome-wide screens and experiments have been instrumental in unraveling cellular and disease properties. Here, we designed and validated a methodology for assembling transcription-activator like effector (TALE) libraries to target any DNA sequence of interest. Our method allows rapid library construction and genome-wide screens.;Combinations of molecular signals such as transcription factors and microRNAs (miRNAs) in cells are a reliable indicator of multi-gene disorders. A system capable of detecting these conditions in-situ may be used as a tool for diagnosis and monitoring of disease. Here, we engineered genetic sensors for combinations of endogenous transcription factors and miRNA. We interfaced our genetic sensors with colorimetric ELISA test strips. Our system allows low-cost, combinatorial, and in-situ reporting of molecular signatures.;Genome editing based on the clustered regularly interspaced short palindromic repeats (CRISPR) system has been expanding at a rapid pace and holds the promise of revolutionizing our ability to probe and edit the genome. We introduce a novel therapeutic technique to target a heterozygous gain-of-function mutation in KRAS. Disrupting the mutation caused the reversal of drug resistance to a MEK small-molecule inhibitor. This technology creates new opportunities for the treatment of a broad range of cancers and genetic diseases.;To summarize, we combine mammalian synthetic biology and genome editing to produce new technologies that advance the field of mammalian synthetic biology bringing us closer to personalized medicine applications in the areas of genetic screening, in-situ sensing, and finally therapeutic targeting.
机译：哺乳动物合成生物学的新兴领域有望彻底改变个性化医学。使用工程遗传电路进行全基因组筛选，检测和报告分子标记以及靶向基因突变可以重新定义个性化药物，其具体应用范围涉及发现到治疗等。全基因组筛选和实验在揭示细胞和疾病方面发挥了重要作用属性。在这里，我们设计并验证了一种组装转录激活子样效应子（TALE）库以靶向任何目标DNA序列的方法。我们的方法允许快速的文库构建和全基因组筛选。;细胞中诸如转录因子和microRNA（miRNA）等分子信号的结合是多基因疾病的可靠指标。能够原位检测这些状况的系统可以用作诊断和监测疾病的工具。在这里，我们设计了用于内源转录因子和miRNA组合的遗传传感器。我们将我们的遗传传感器与比色ELISA试纸对接。我们的系统允许进行低成本，组合和就地报告分子特征的报告;基于簇状规则间隔的短回文重复序列（CRISPR）的基因组编辑正在迅速扩展，并有望彻底改变我们的能力。探测和编辑基因组。我们介绍一种新型的治疗技术，以针对KRAS中的杂合功能获得性突变。破坏突变导致对MEK小分子抑制剂的耐药性逆转。这项技术为广泛的癌症和遗传疾病的治疗创造了新的机会。总而言之，我们结合了哺乳动物合成生物学和基因组编辑技术，产生了促进哺乳动物合成生物学领域发展的新技术，使我们更接近于个性化医学应用。基因筛选，原位感测和治疗靶向领域。

著录项

  • 作者

    Ehrhardt, Kristina Jeanette.;

  • 作者单位

    The University of Texas at Dallas.;

  • 授予单位 The University of Texas at Dallas.;
  • 学科 Biomedical engineering.;Genetics.
  • 学位 Ph.D.
  • 年度 2016
  • 页码 93 p.
  • 总页数 93
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 康复医学;
  • 关键词

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