sirolimus and alpha-sarcin

Protein splicing technology harnesses the ability of inteins to ligate protein fragments, forming a mature protein. This report describes our effort to engineer rapamycin-dependent protein splicing of a ribotoxin, called α-sarcin. Engineering this system required the investigation of important splicing parameters, including extein context and splicing temperature. We show α-sarcin splicing is dependent on rapamycin, is inducible with rapid kinetics, and triggers apoptosis in HeLa cells. These findings establish a proof-of-concept for a conditional cell ablation strategy.

Topics: Apoptosis; Cell Line, Tumor; Endoribonucleases; Fungal Proteins; Green Fluorescent Proteins; HeLa Cells; Humans; Inteins; Protein Engineering; Protein Folding; Protein Splicing; Sirolimus