Inducible Degradation of Target Proteins through a Tractable Affinity-Directed Protein Missile System

Luke M. Simpson, Thomas J. Macartney, Alice Nardin, Luke J. Fulcher, Sascha Röth, Andrea Testa, Chiara Maniaci, Alessio Ciulli, Ian G. Ganley, Gopal P. Sapkota (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

36 Citations (Scopus)
90 Downloads (Pure)


The affinity-directed protein missile (AdPROM) system utilizes specific polypeptide binders of intracellular proteins of interest (POIs) conjugated to an E3 ubiquitin ligase moiety to enable targeted proteolysis of the POI. However, a chemically tuneable AdPROM system is more desirable. Here, we use Halo-tag/VHL-recruiting proteolysis-targeting chimera (HaloPROTAC) technology to develop a ligand-inducible AdPROM (L-AdPROM) system. When we express an L-AdPROM construct consisting of an anti-GFP nanobody conjugated to the Halo-tag, we achieve robust degradation of GFP-tagged POIs only upon treatment of cells with the HaloPROTAC. For GFP-tagged POIs, ULK1, FAM83D, and SGK3 were knocked in with a GFP-tag using CRISPR/Cas9. By substituting the anti-GFP nanobody for a monobody that binds H- and K-RAS, we achieve robust degradation of unmodified endogenous RAS proteins only in the presence of the HaloPROTAC. Through substitution of the polypeptide binder, the highly versatile L-AdPROM system is useful for the inducible degradation of potentially any intracellular POI.

Original languageEnglish
Pages (from-to)1164-1180.e5
Number of pages17
JournalCell Chemical Biology
Issue number9
Early online date14 Jul 2020
Publication statusPublished - 17 Sept 2020


  • AdPROM
  • HaloPROTAC
  • nanobody
  • monobody
  • targeted proteolysis
  • ULK1
  • FAM83D
  • SGK3
  • RAS

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry


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