Engineered PROTAC-CID Systems for Mammalian Inducible Gene Regulation

Gene regulation via chemically induced dimerization (CID) is useful for biomedical research. However, the number, type, versatility, and in vivo applications of CID tools remain limited. Here, we demonstrate the development of proteolysis-targeting chimera-based scalable CID (PROTAC-CID) platforms by systematically engineering the available PROTAC systems for inducible gene regulation and gene editing. Further, we show orthogonal PROTAC-CIDs that can fine-tune gene expression at gradient levels or multiplex biological signals with different logic gating operations. Coupling the PROTAC-CID platform with genetic circuits, we achieve digitally inducible expression of DNA recombinases, base-and prime-editors for transient genome manipulation. Finally, we package a compact PROTAC-CID system into adeno-associated viral vectors for inducible and reversible gene activation in vivo. This work provides a versatile molecular toolbox that expands the scope of chemically inducible gene regulation in human cells and mice.

Automated summary

This study developed a scalable chemically inducible dimerization platform (PROTAC-CID) based on proteolysis-targeting chimera, which enables inducible gene regulation and gene editing. It can fine-tune gene expression at gradient levels or multiplex biological signals. By coupling it with genetic circuits, digitally inducible expression of DNA recombinases and base-and prime-editors for transient genome manipulation are achieved. Furthermore, a compact PROTAC-CID system is packaged into adeno-associated viral vectors for inducible and reversible gene activation in vivo. This work provides a versatile molecular toolbox for chemically inducible gene regulation in human cells and mice.