Rapid and specific degradation of endogenous proteins in mouse models using auxin-inducible degrons

Macdonald, L. A.; Taylor, G. C. A.; Brisbane, J. M.; Christodoulou, E.; Scott, L.; Von Kriegsheim, A.; Rossant, J. J.; Gu, B.; Wood, A. J.

doi:10.1101/2022.01.13.476100

bioRxiv · 10.1101/2022.01.13.476100

Rapid and specific degradation of endogenous proteins in mouse models using auxin-inducible degrons

Macdonald, L. A.·Taylor, G. C. A.·Brisbane, J. M.·Christodoulou, E.·Scott, L.·Von Kriegsheim, A.·Rossant, J. J.·Gu, B.·Wood, A. J.

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Abstract

Auxin-inducible degrons are a chemical genetic tool for targeted protein degradation and are widely used to study protein function in cultured mammalian cells. Here, we develop CRISPR-engineered mouse lines that enable rapid and highly specific degradation of tagged endogenous proteins in vivo. Most but not all cell types are competent for degradation. Using mouse genetics, we show that degradation kinetics depend upon the dose of the tagged protein, ligand, and the E3 ligase subunit Tir1. Rapid degradation of condensin I and condensin II - two essential regulators of mitotic chromosome structure - revealed that both complexes are individually required for cell division in precursor lymphocytes, but not in their differentiated peripheral lymphocyte derivatives. This generalisable approach provides unprecedented temporal control over the dose of endogenous proteins in mouse models, with implications for studying essential biological pathways and modelling drug activity in mammalian tissues. HighlightsO_LIAuxin-inducible degradation of endogenously tagged proteins in living mice and a range of primary cells. C_LIO_LIMost but not all cell types are competent for degradation C_LIO_LIDosage of the tagged protein, E3 ligase substrate receptor and ligand can all determine degradation kinetics C_LIO_LIRapid degradation of condensin subunits in lymphocytes reveals stage-specific requirements during cell division C_LI

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Macdonald, L. A., Taylor, G. C. A., Brisbane, J. M., Christodoulou, E., Scott, L., Von Kriegsheim, A., Rossant, J. J., Gu, B., Wood, A. J.. 2022-01-13. Rapid and specific degradation of endogenous proteins in mouse models using auxin-inducible degrons. https://doi.org/10.1101/2022.01.13.476100

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