bioRxivBioengineering
A super sensitive auxin-inducible degron system with an engineered auxin-TIR1 pair
K. Nishimura, R. Yamada, S. Hagihara, R. Iwasaki, N. Uchida, T. Kamura, K. Takahashi, K. U. Torii, T. Fukagawa
Abstract
Auxin-Inducible Degron (AID) technology enables conditional depletion of targeted proteins. However, the applicability of the AID in vertebrate cells has been limited due to cytotoxicity caused by high auxin concentrations. Here, we establish an improved AID system using an engineered orthogonal auxin-TIR1 pair, which exhibits over 1,000 times stronger binding. With ~1,000-fold less auxin concentration, we achieved to generate the AID-based knockout cells in various human and mouse cell lines in a single transfection.
- organism: human
- organism: mouse
- research focus: protein folding & dynamics
- research focus: genome assembly
- approach: experimental assay
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