Engineered PROTAC-CID Systems for Mammalian Inducible Gene Regulation

Authors

Dacheng Ma
Qichen Yuan
Fei Peng
Victor Paredes
Hongzhi Zeng
Emmanuel C Osikpa
Qiaochu Yang
Advaith Peddi
Anika Patel
Megan S Liu
Zheng Sun
Xue Gao

Publication Date

1-25-2023

Journal

Journal of the American Chemical Society

DOI

10.1021/jacs.2c09129

PMID

36626587

PMCID

PMC10162582

PubMedCentral® Posted Date

5-5-2023

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

yes

Keywords

Humans, Mice, Animals, Dimerization, DNA, Recombinases, Gene Editing, Genome, CRISPR-Cas Systems, Mammals, PROTAC, CID, inducible gene regulation, mammal, genome editing, AAV delivery

Abstract

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.

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