Harnessing nanobodies for target protein degradation through the Affinity-directed PROtein Missile (AdPROM) system

Bill Carton, Sascha Röth, Thomas J. Macartney, Gopal P. Sapkota (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)


Targeted protein degradation (TPD) is a useful approach in dissecting protein function and therapeutics. Technologies such as RNA interference or gene knockout that are routinely used rely on protein turnover. However, RNA interference takes a long time to deplete target proteins and is not suitable for long-lived proteins, while a genetic knockout is irreversible, takes a long time to achieve and is not suitable for essential genes. TPD has the potential to overcome the limitations of RNA interference and gene editing approaches. We have established the Affinity directed PROtein Missile (AdPROM) system, which harnesses nanobodies or binders of target proteins to redirect E3 ubiquitin ligase activity to the target protein to induce TPD through the ubiquitin proteasome system. Here we provide a step-by-step protocol for using the AdPROM system for targeted proteolysis of endogenously GFP-tagged K-RAS through an anti-GFP nanobody. This protocol can be amended to target a wide range of different proteins of interest (POIs) either by replacing the anti-GFP nanobody with a nanobody recognising the POI or by endogenously tagging the POI with GFP through CRISPR/Cas9 genome editing.

Original languageEnglish
Title of host publicationTargeted Protein Degradation
EditorsGeorge L. Burslem
Place of PublicationCambridge, MA
PublisherAcademic Press Inc.
Number of pages19
ISBN (Print)9780323992008
Publication statusPublished - 2023

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • AdPROM
  • E3
  • Proteasome
  • Target protein degradation
  • TPD
  • Ubiquitin
  • Ubiquitination

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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