2. Academic Validation
  3. A CRISPR activation screen identifies FBXO22 supporting targeted protein degradation

A CRISPR activation screen identifies FBXO22 supporting targeted protein degradation

  • Nat Chem Biol. 2024 Jul 4. doi: 10.1038/s41589-024-01655-9.
Ananya A Basu 1 2,  Chenlu Zhang 1,  Isabella A Riha 1,  Assa Magassa 1 2,  Miguel A Campos 1 2,  Alana G Caldwell 1 3,  Felicia Ko 1,  Xiaoyu Zhang 4 5 6 7 8
Affiliations

Affiliations

  • 1 Department of Chemistry, Northwestern University, Evanston, IL, USA.
  • 2 Chemistry of Life Processes Institute, Northwestern University, Evanston, IL, USA.
  • 3 Interdisciplinary Biological Sciences Program, Northwestern University, Evanston, IL, USA.
  • 4 Department of Chemistry, Northwestern University, Evanston, IL, USA. zhang@northwestern.edu.
  • 5 Chemistry of Life Processes Institute, Northwestern University, Evanston, IL, USA. zhang@northwestern.edu.
  • 6 Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, IL, USA. zhang@northwestern.edu.
  • 7 Center for Human Immunobiology, Northwestern University, Chicago, IL, USA. zhang@northwestern.edu.
  • 8 International Institute for Nanotechnology, Northwestern University, Evanston, IL, USA. zhang@northwestern.edu.
PMID: 38965383 DOI: 10.1038/s41589-024-01655-9
Abstract

Targeted protein degradation (TPD) represents a potent chemical biology paradigm that leverages the cellular degradation machinery to pharmacologically eliminate specific proteins of interest. Although multiple E3 Ligases have been discovered to facilitate TPD, there exists a compelling requirement to diversify the pool of E3 Ligases available for such applications. Here we describe a clustered regularly interspaced short palindromic repeats (CRISPR)-based transcriptional activation screen focused on human E3 Ligases, with the goal of identifying E3 Ligases that can facilitate heterobifunctional compound-mediated target degradation. Through this approach, we identified a candidate proteolysis-targeting chimera (PROTAC), 22-SLF, that induces the degradation of FK506-binding protein 12 when the transcription of FBXO22 gene is activated. Subsequent mechanistic investigations revealed that 22-SLF interacts with C227 and/or C228 in F-box protein 22 (FBXO22) to achieve target degradation. Lastly, we demonstrated the versatility of FBXO22-based PROTACs by effectively degrading additional endogenous proteins, including bromodomain-containing protein 4 and the echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase fusion protein.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-163807
    PROTAC Degrader for FKBP12