idTAG enables programmable and physiologically faithful control of endogenous protein degradation
- 1Department of Hematology, Medical Research Institute, Frontier Science Center for Immunology and Metabolism, Zhongnan Hospital of Wuhan University, Wuhan Univer
Abstract
Rapid and precise depletion of endogenous proteins is essential for dissecting gene function, yet existing degron technologies often require direct modification of endogenous coding sequences, which can compromise protein expression and limit their applicability. Here, we develop idTAG, an inducible degron platform that decouples protein expression from protein degradation by integrating Tet-On 3G-mediated transcriptional control with dTAG-mediated degradation. Systematic characterization reveals that conventional degron knock-in frequently causes unintended reduction of target protein expression at the mRNA level, representing an underappreciated limitation of current approaches. In contrast, idTAG enables efficient generation of degron cell lines while maintaining physiological protein abundance and allows programmable control of degradation kinetics through independent regulation of protein synthesis and degradation. Applying idTAG to RNA-binding proteins, we demonstrate that acute depletion of FUS has minimal effects on global RNA polymerase II transcription, whereas acute loss of SRSF5 selectively disrupts transcription termination at a subset of genes. Thus, idTAG provides a versatile platform for physiological and temporally controlled interrogation of endogenous protein functions.
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BibTeX
@misc{san2026idtag,
title = {{idTAG enables programmable and physiologically faithful control of endogenous protein degradation}},
author = {M. San and J. Peng and F. Zhao and R. Xiao},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.18.752548},
url = {https://www.biorxiv.org/content/10.64898/2026.09.18.752548v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - idTAG enables programmable and physiologically faithful control of endogenous protein degradation
AU - M. San
AU - J. Peng
AU - F. Zhao
AU - R. Xiao
PY - 2026
DA - 2026/09/18
PB - bioRxiv
DO - 10.64898/2026.09.18.752548
UR - https://www.biorxiv.org/content/10.64898/2026.09.18.752548v1
AB - Rapid and precise depletion of endogenous proteins is essential for dissecting gene function, yet existing degron technologies often require direct modification of endogenous coding sequences, which can compromise protein expression and limit their applicability. Here, we develop idTAG, an inducible degron platform that decouples protein expression from protein degradation by integrating Tet-On 3G-mediated transcriptional control with dTAG-mediated degradation. Systematic characterization reveals that conventional degron knock-in frequently causes unintended reduction of target protein expression at the mRNA level, representing an underappreciated limitation of current approaches. In contrast, idTAG enables efficient generation of degron cell lines while maintaining physiological protein abundance and allows programmable control of degradation kinetics through independent regulation of protein synthesis and degradation. Applying idTAG to RNA-binding proteins, we demonstrate that acute depletion of FUS has minimal effects on global RNA polymerase II transcription, whereas acute loss of SRSF5 selectively disrupts transcription termination at a subset of genes. Thus, idTAG provides a versatile platform for physiological and temporally controlled interrogation of endogenous protein functions.
ER -