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bioRxiv · 10.1101/2024.06.04.597287

The AID2 system offers a potent tool for rapid, reversible, or sustained degradation of essential proteins in live mice

Sladky, V. C.·Strong, M. A.·Tapias-Gomez, D.·Jewett, C. E.·Drown, C. G.·Scott, P. M.·Holland, A. J.
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Abstract

Studying essential genes required for dynamic processes in live mice is challenging as genetic perturbations are irreversible and limited by slow protein depletion kinetics. The first-generation auxin-inducible-degron (AID) system is a powerful tool for analyzing inducible protein loss in cultured cells. However, auxin administration is toxic to mice, preventing its long-term use in animals. Here, we use an optimized second-generation AID system to achieve the conditional and reversible loss of the essential centrosomal protein CEP192 in live mice. We show that the auxin derivative 5-Ph-IAA is well tolerated over two weeks and drives near-complete CEP192-mAID degradation in less than one hour in vivo. Prolonged CEP192 loss led to cell division failure and cell death in proliferative tissues. Thus, the second-generation AID system is well suited for rapid and/or sustained protein depletion in live mice, offering a valuable new tool for interrogating protein function in vivo.

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Sladky, V. C., Strong, M. A., Tapias-Gomez, D., Jewett, C. E., Drown, C. G., Scott, P. M., Holland, A. J.. 2024-06-05. The AID2 system offers a potent tool for rapid, reversible, or sustained degradation of essential proteins in live mice. https://doi.org/10.1101/2024.06.04.597287

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