Genetically Encoded Biosensor for Fluorescence Lifetime Imaging of PTEN Dynamics in the Intact Brain

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2025 Feb 20

PMID 39979596

Authors

Tomer Kagan

Matan Gabay

Aasha Meenakshisundaram

Yossi Levi

Sharbel Eid

Nikol Malchenko

Maya Maman

Anat Nitzan

Luca Ravotto

Ronen Zaidel-Bar

Britta Johanna Eickholt

Maayan Gal

Tal Laviv

Affiliations

Soon will be listed here.

Abstract

The phosphatase and tensin homolog (PTEN) is a vital protein that maintains an inhibitory brake for cellular proliferation and growth. Accordingly, PTEN loss-of-function mutations are associated with a broad spectrum of human pathologies. Despite its importance, there is currently no method to directly monitor PTEN activity with cellular specificity within intact biological systems. Here we describe the development of a FRET-based biosensor using PTEN conformation as a proxy for the PTEN activity state, for two-photon fluorescence lifetime imaging microscopy. We identify a point mutation that allows the monitoring of PTEN activity with minimal interference to endogenous PTEN signaling. We demonstrate imaging of PTEN activity in cell lines, intact Caenorhabditis elegans and in the mouse brain. Finally, we develop a red-shifted sensor variant that allows us to identify cell-type-specific PTEN activity in excitatory and inhibitory cortical cells. In summary, our approach enables dynamic imaging of PTEN activity in vivo with unprecedented spatial and temporal resolution.

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