An Analog-Sensitive Tau Tubulin Kinase 2 Enables Acute Chemical-Genetic Control of Primary Cilia
Abstract
Kinases regulate a broad spectrum of cellular processes, including the formation of primary cilia, microtubule-based organelles that protrude from the cell surface and play important roles in development and disease. TTBK2 is a key ciliary kinase that controls primary cilium assembly and maintenance. However, highly selective small-molecule inhibitors of TTBK2 are still lacking, representing a major bottleneck for investigating the temporal requirements and dynamics of TTBK2-dependent processes in cilia. Here, we report the first analog-sensitive variant of TTBK2, enabling rapid, reversible, and highly specific control of TTBK2 kinase activity in ciliogenesis-relevant hTERT-RPE1 cells. Using this system, we define the kinetics of CEP164 phosphorylation, cilium shortening and reassembly, and IFT88 retention at the ciliary base following acute TTBK2 inhibition and subsequent inhibitor washout. Together, our findings provide new insight into TTBK2-dependent pathways and establish analog-sensitive TTBK2 as a powerful tool for investigating the dynamics of TTBK2-regulated processes with high temporal resolution.
Related
- Engineering Bacteria and Their Vesicles for Therapy: Integrating Native Functions, Genetic Control, and Chemical Biology
- An engineered α-1,3-glucanase-derived probe enables sensitive detection of α-1,3-glucan and reveals its soluble release by Aspergillus
- Compression Sequencing enables ultra-sensitive and scalable scRNA-seq
- Gel-Confined Rolling-Circle Amplification Enables Sensitive Single-Cell Proteoform Analysis
- Decoder-FFPE-seq enables sensitive, genome-wide spatial transcriptomics of archival tissues at single-cell resolution
BibTeX
@misc{lacigova2026analogsensitive,
title = {{An Analog-Sensitive Tau Tubulin Kinase 2 Enables Acute Chemical-Genetic Control of Primary Cilia}},
author = {A. Lacigova and L. Cajanek},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.09.757601},
url = {https://www.biorxiv.org/content/10.64898/2026.10.09.757601v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - An Analog-Sensitive Tau Tubulin Kinase 2 Enables Acute Chemical-Genetic Control of Primary Cilia
AU - A. Lacigova
AU - L. Cajanek
PY - 2026
DA - 2026/10/09
PB - bioRxiv
DO - 10.64898/2026.10.09.757601
UR - https://www.biorxiv.org/content/10.64898/2026.10.09.757601v1
AB - Kinases regulate a broad spectrum of cellular processes, including the formation of primary cilia, microtubule-based organelles that protrude from the cell surface and play important roles in development and disease. TTBK2 is a key ciliary kinase that controls primary cilium assembly and maintenance. However, highly selective small-molecule inhibitors of TTBK2 are still lacking, representing a major bottleneck for investigating the temporal requirements and dynamics of TTBK2-dependent processes in cilia. Here, we report the first analog-sensitive variant of TTBK2, enabling rapid, reversible, and highly specific control of TTBK2 kinase activity in ciliogenesis-relevant hTERT-RPE1 cells. Using this system, we define the kinetics of CEP164 phosphorylation, cilium shortening and reassembly, and IFT88 retention at the ciliary base following acute TTBK2 inhibition and subsequent inhibitor washout. Together, our findings provide new insight into TTBK2-dependent pathways and establish analog-sensitive TTBK2 as a powerful tool for investigating the dynamics of TTBK2-regulated processes with high temporal resolution.
ER -