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. 2016 Jan 20;6(1):150230. doi: 10.1098/rsob.150230

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© 2016 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 1. — Schematic of the conditional knockout strategy and its adaption for the AID-CENP-T cell line. (a) Doxycycline regulated shut-down of CENP-T or CENP-W gene expression. (b) Combining the doxycycline regulated shut-down of gene expression with the auxin-inducible degron (AID) system. (c) A schematic of the auxin-induced degradation system applied in eukaryotes. The SCF E3 ubiquitin ligase consists of 4 units: Rbx1, Cul-1, Skp1 and F-box containing protein, TIR1. Auxin hormone (such as IAA) binding to the ectopically expressed TIR1 receptor promotes contacts with target proteins fused to an AID tag. The SCF-TIR1 E3 ligase can then polyubiquitinate the AID tag, promoting degradation of the substrate by the 26S proteasome.