iTAG an optimized IMiD-induced degron for targeted protein degradation in human and murine cells

Habib Bouguenina; Stephanos Nicolaou; Yann-Vaï Le Bihan; Elizabeth A Bowling; Cheyenne Calderon; John J Caldwell; Brinley Harrington; Angela Hayes; P Craig McAndrew; Costas Mitsopoulos; Fernando Jr Sialana; Andrea Scarpino; Mark Stubbs; Arjun Thapaliya; Siddhartha Tyagi; Hannah Z Wang; Francesca Wood; Rosemary Burke; Florence Raynaud; Jyoti Choudhary; Rob L M van Montfort; Amine Sadok; Thomas F Westbrook; Ian Collins; Rajesh Chopra

doi:10.1016/j.isci.2023.107059

iTAG an optimized IMiD-induced degron for targeted protein degradation in human and murine cells

iScience. 2023 Jun 7;26(7):107059. doi: 10.1016/j.isci.2023.107059. eCollection 2023 Jul 21.

Authors

Habib Bouguenina¹, Stephanos Nicolaou¹, Yann-Vaï Le Bihan¹, Elizabeth A Bowling², Cheyenne Calderon², John J Caldwell¹, Brinley Harrington², Angela Hayes¹, P Craig McAndrew¹, Costas Mitsopoulos¹, Fernando Jr Sialana^{1

3}, Andrea Scarpino¹, Mark Stubbs¹, Arjun Thapaliya¹, Siddhartha Tyagi², Hannah Z Wang¹, Francesca Wood¹, Rosemary Burke¹, Florence Raynaud¹, Jyoti Choudhary³, Rob L M van Montfort¹, Amine Sadok¹, Thomas F Westbrook², Ian Collins¹, Rajesh Chopra¹

Affiliations

¹ Centre for Cancer Drug Discovery, the Institute of Cancer Research, 15 Cotswold Road, Sutton, London SM2 5NG, UK.
² Therapeutic Innovation Centre (THINC), Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
³ Functional Proteomics Group, The Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, UK.

Abstract

To address the limitation associated with degron based systems, we have developed iTAG, a synthetic tag based on IMiDs/CELMoDs mechanism of action that improves and addresses the limitations of both PROTAC and previous IMiDs/CeLMoDs based tags. Using structural and sequence analysis, we systematically explored native and chimeric degron containing domains (DCDs) and evaluated their ability to induce degradation. We identified the optimal chimeric iTAG(DCD23 60aa) that elicits robust degradation of targets across cell types and subcellular localizations without exhibiting the well documented "hook effect" of PROTAC-based systems. We showed that iTAG can also induce target degradation by murine CRBN and enabled the exploration of natural neo-substrates that can be degraded by murine CRBN. Hence, the iTAG system constitutes a versatile tool to degrade targets across the human and murine proteome.

Keywords: Biochemistry; Biological sciences; Cell biology; Oncology.