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

Carnegie Mellon University, Pittsburgh PA

Investigators

Abstract

0748062 Zappe The 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'.