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)

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)
200 Downloads (Pure)

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

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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