Target Identification

At the cellular level, important research strategies in elucidating gene and protein function include knock-down techniques (for example, RNAi and CALI), developmental studies in wild-type and mutant organisms, and protein localization and protein interaction studies. Microscopy is an essential tool in these target identification strategies and in monitoring the effects of investigational compound on these targets.

Microscopy is used for micromanipulation when transfecting cells with knock-down molecules and verifying uptake and localization within cells and tissues using GFP tags. It is also important in visualizing directly the effects of gene knock-down and gene mutation in both developing organisms and in individual cells using time-lapse imaging. Microscopy is also essential for observing the downstream effects of gene and protein manipulation at the molecular level. Protein / protein interactions, protein dynamics and localization can all be observed using fluorescent probes and technique such as FRET/BRET, FRAP and TIRF.

Key light microscopy techniques in target identification include fluorescence, confocal, multidimensional imaging, spectral imaging, FRET/BRET, FRAP, FLIM, and TIRF, photoactivatable GFP, kaede, and quantum dots imaging.