CAREER: Automated MEMS-based Drosophila Embryo Injection Technologies for High-Throughput Functional Genomics Screens

果蝇是人类生物学的重要模式生物。 除了D.除了黑腹果蝇外，其他11种果蝇的基因组序列在过去两年中也已公开。 这些基因组序列为研究基因的功能及其在发育和疾病中的意义带来了前所未有的机会。 基因功能通常是从生物体遗传组成的特定扰动和随后对生物体影响的分析中推断出来的。 在过去已经建立了两个强大的方法，在果蝇这样的分析：永久转座遗传元件的遗传转化和瞬时特异性基因沉默通过RNA干扰（RNAi）。 这两种方法都需要在胚胎发育的最早阶段分别可靠和快速地注射DNA和双链RNA（dsRNA）。在NSF CAREER项目的框架内，将开发两种自动化的基于MEMS的果蝇胚胎注射系统，称为“搜索和注射”和“饲料和注射”，以实现对果蝇胚胎基因功能的高通量筛选。 “搜索和注射”将支持RNAi实验;“饲料和注射”将支持遗传转化实验。 与最先进的手动进样程序相比，自动进样技术将使实验通量增加约20倍。 开发超出原理证明的强大技术将使其能够在果蝇研究界传播和广泛使用。 在NSF CAREER奖下开发的注射系统将与果蝇胚胎的高通量共聚焦成像系统以及用于可靠自动识别基因沉默表型的图像分析软件相补充。 这些新工具的应用将使人们更好地了解人类发育和疾病的分子机制，预计将对新疗法和改善公共卫生状况产生重大影响。 拟议的研究将推进有关MEMS器件的设计，制造，封装和应用的基础工程知识。 所产生的知识可以帮助创建用于自动处理DNA、RNA、其他生化试剂、细胞、卵母细胞、胚胎以及微米和纳米颗粒的系统，在生物研究、生物技术、药物发现、高通量筛选和医学诊断中具有广泛的应用。和本科生在科学生涯的早期，并教育一个新的工程师谁可以创造性地确定和解决生物学和医学的技术需求。外联活动包括为少数民族学生举办的为期十周的本科生暑期研究项目，为高中生举办的为期六周的暑期研究项目，以及为中学生举办的周末研讨会。 研究也直接整合在两个新的跨学科类：“生物MEMS和生物医学纳米技术”和“干细胞工程”。

0748062ZappeThe fruit fly Drosophila serves as an important model organism for human biology. In addition to the genome sequence of D. melanogaster, the genome sequences of 11 other Drosophila species have been made publicly available over the past two years. These genome sequences have brought unprecedented opportunities to study functions of genes and their implications in development and disease. Gene functions are often inferred from specific perturbation of the genetic makeup of an organism and subsequent analysis of the effects on the organism. Two powerful methods have been established in the past for such analysis in Drosophila: permanent genetic transformation with transposable genetic elements and transient specific gene silencing through RNA interference (RNAi). Both methods require reliable and rapid injection of DNA and doublestranded RNA (dsRNA), respectively, at the earliest stages of embryonic development.Within the frame of this NSF CAREER project, two automated, MEMS-based Drosophila embryo injection systems termed 'Search and Inject' and 'Feed and Inject' will be developed to enable high-throughput screens for gene functions in Drosophila embryos. 'Search and Inject' will support RNAi experiments; 'Feed and Inject' will support genetic transformation experiments. The automated injection technologies will lead to an approximately 20-fold increase in experimental throughput compared to state-of-the-art manual injection procedures. Development of robust technologies beyond proof of principle will enable their dissemination and widespread use within the Drosophila research community. The injection systems developed under the NSF CAREER award will be complemented with a system for high-throughput, confocal imaging of Drosophila embryos as well as image analysis software for reliable, automated recognition of phenotypes due to gene silencing. Application of these new tools will lead to a better understanding of molecular mechanisms of development and disease in humans, with expected significant impact on new therapies and improvement of the state of public health. The proposed research will advance fundamental engineering knowledge regarding design, fabrication, packaging, and application of MEMS devices. The generated knowledge can help create systems for automated handling of DNA, RNA, other biochemical reagents, cells, oocytes, embryos, as well as micro- and nanoparticles, with widespread applications in biological research, biotechnology, drug discovery, high-throughput screening, and medical diagnostics.Educational and outreach activities are designed to interest middle school, high school, and undergraduate students early on in a career in science and to educate a new breed of engineers who can creatively identify and address technological needs in biology and medicine. Outreach activities include a ten-week-long undergraduate summer research program for minority students, a six-week-long summer research program for high school students, and a weekend workshop for middle school students. Research is also directly integrated in two new, interdisciplinary classes: 'BioMEMS and Biomedical Nanotechnology' and 'Stem Cell Engineering'.