A dual-color fluorescence-based platform to identify selective inhibitors of Akt signaling.

A dual-color fluorescence-based platform to identify selective inhibitors of Akt signaling.

Blog Article

BACKGROUND: Inhibition of Akt signaling is considered one of the most promising therapeutic strategies for many cancers.However, rational target-orientated approaches to cell based drug screens for anti-cancer agents have historically been compromised by the Chargers notorious absence of suitable control cells.METHODOLOGY/PRINCIPAL FINDINGS: In order to address this fundamental problem, we have developed BaFiso, a live-cell screening platform to identify specific inhibitors of this pathway.BaFiso relies on the co-culture of isogenic cell lines that have been engineered to sustain interleukin-3 independent survival of the parental Ba/F3 cells, and that are individually tagged with different fluorescent proteins.

Whilst in the first of these two lines cell survival in the absence of IL-3 is dependent on the expression of activated Akt, the cells expressing constitutively-activated Stat5 signaling display IL-3 independent growth and survival in an Akt-independent manner.Small molecules can then be screened in these lines to identify inhibitors that rescue IL-3 dependence.CONCLUSIONS/SIGNIFICANCE: BaFiso measures differential cell survival using multiparametric live cell imaging and permits selective inhibitors of Akt signaling to DRESSES be identified.BaFiso is a platform technology suitable for the identification of small molecule inhibitors of IL-3 mediated survival signaling.