Zebrafish has emerged as a powerful in vivo platform for phenotype-based drug screens and chemical genetic analysis. Here, we demonstrate a simple, practical method for large-scale screening of small molecules using zebrafish embryos.
1) Zebrafish Egg Collection
2) Arraying Embryos in 96-well Plates
3) Transfer of Small Molecule Library
While compound transfer can be automated with robotic transfer methods, we will describe the manual transfer method.
4) Screening for Effects of Small Molecules by Visual Inspection of Phenotypes
When planning a zebrafish-based chemical screen, particular attention must be paid to the robustness of the phenotype under consideration and the background rate of such phenotype. This is particularly important for screens for chemical suppressors of an induced phenotype. For example, for a phenotype caused by heat-shock induction of a transgene, the condition that induces the phenotype reproducibly must be precisely mapped out prior to initiating the screen to avoid unacceptably high false positive rates. With careful planning, zebrafish chemical screens, which require modest capital investment, can lead to the discovery of novel modulator of cell signaling and physiology, as well as lead compound to reverse models of human diseases. Lastly, by analogy to the classic forward genetic screens, zebrafish-based chemical screens can be valuable for chemical genetic dissection of fundamental biological process.
The authors have nothing to disclose.