Microsystems for controlled genetic perturbation of live Drosophila embryos: RNA interference, development robustness and drug screening

作者：Fakhoury Jean R; Sisson John C; Zhang Xiaojing^*

来源：Microfluidics and Nanofluidics, 2009, 6(3): 299-313.

DOI：10.1007/s10404-009-0405-x

• Drosophila embryo
• RNA interference
• Microinjection
• Drug screening
• Microfluidic self assembly
• Perturbation

A combination of genetic regulation and microenvironmental perturbation might prove especially useful for identifying the fundamental mechanisms responsible for development biology and critical bio-chemical networks. This could transform our understanding of genomics, proteomics, and drug discovery and lead to personalized drugs and molecular diagnostics for improved healthcare. With the complete sequencing of the Drosophila melanogaster, there has been a growing interest in correlating its genetics to human disease. The Drosophila embryo is an excellent whole-animal model, and is ideal for high throughput analysis. This review highlights research on the recent development of miniature tools to study critical cellular processes in Drosophila embryogenesis. First, we discuss the use of micro-injectors to conduct chip-level RNA interference experiments on self-assembled Drosophila embryos in order to determine how genes contribute to the growth and function of a developing organism. Second, we review recent progress on perturbing the microenvironment as a complementary approach to perturbing the genetic components of the development network. We designed a bio-compatible microfluidic device that automatically positions live embryos within a fluid-filled imaging channel and exposes embryos to constant or biphasic temperature or different drug concentrations. We have demonstrated that a thermal gradient applied across live Drosophila embryos results in asynchronous cellularization. Additionally, preliminary drug screening experiments were carried out on live Drosophila embryos in both 96-well plates and microfluidic channels. Time-lapse differential interference contrast microscopic images were taken to show the effects of colchicine (399.40 Da) and cytochalasin D (507.62 Da) on cellularization.

• 出版日期2009-3