Antibody RING-Mediated Destruction of Endogenous Proteins

Adel F. M. Ibrahim, Linnan Shen, Michael H. Tatham, David Dickerson, Alan R. Prescott, Naima Abidi, Dimitris P. Xirodimas, Ronald T. Hay (Lead / Corresponding author)

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Abstract

To understand gene function, the encoding DNA or mRNA transcript can be manipulated and the consequences observed. However, these approaches do not have a direct effect on the protein product of the gene, which is either permanently abrogated or depleted at a rate defined by the half-life of the protein. We therefore developed a single-component system that could induce the rapid degradation of the specific endogenous protein itself. A construct combining the RING domain of ubiquitin E3 ligase RNF4 with a protein-specific camelid nanobody mediates target destruction by the ubiquitin proteasome system, a process we describe as antibody RING-mediated destruction (ARMeD). The technique is highly specific because we observed no off-target protein destruction. Furthermore, bacterially produced nanobody-RING fusion proteins electroporated into cells induce degradation of target within minutes. With increasing availability of protein-specific nanobodies, this method will allow rapid and specific degradation of a wide range of endogenous proteins.

Original languageEnglish
Pages (from-to)155-166
Number of pages12
JournalMolecular Cell
Volume79
Issue number1
Early online date25 May 2020
DOIs
Publication statusPublished - 2 Jul 2020

Keywords

  • ARMeD
  • E3 ligase
  • nanobody-RING fusion
  • proteasome
  • protein degradation
  • ubiquitin

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