# Generation and characterization of a tamoxifen-inducible, Cre driver rat for transgene expression in microglia

## TL;DR

This paper introduces a new rat model that allows for tamoxifen-inducible genetic manipulation of microglia, enabling studies of their role in the brain and immune system.

## Contribution

The first tamoxifen-inducible Cre driver rat for microglia-specific transgene expression in rats.

## Key findings

- The Cx3cr1-CreERT2 rat model shows microglial-specific Cre expression confirmed by mCherry reporter colocalization with Iba1.
- Flow cytometry revealed time- and Cre-dependent recombination in Cx3cr1+ cells across spleen, blood, and brain.
- The model maintains normal cognitive behavior compared to wildtype controls.

## Abstract

Microglia are the resident immune cells of the central nervous system (CNS) and display diverse functions under both physiological and pathological conditions. The past decade has seen burgeoning interest in microglia function, with a variety of transgenic tools developed for specific genetic manipulation of microglia in various injury, disease, and developmental models. Although many of these models have been developed in mice, the ability to manipulate microglia in rats provides additional advantages to studying microglial function in the brain especially related to complex behavior. Using BAC transgenesis, our lab created a transgenic rat (Cx3cr1-CreERT2) that expresses a tamoxifen inducible Cre recombinase (CreERT2) under control of the microglial/macrophage specific fractalkine C-X3-C Motif Chemokine Receptor 1 (Cx3cr1) promoter. In mice, CreERT2 and other transgenes have been expressed in microglia using the Cx3cr1 promoter, however, this is the first demonstration in rats. Importantly, these rats exhibit similar cognitive behaviors compared to their wildtype (WT) controls. Microglial specificity of inducible Cre expression was confirmed by breeding the novel Cx3cr1-CreERT2+/− rat with a previously reported double floxed inverse open reading frame (DIO)-mCherry+/− reporter rat to show tamoxifen inducible mCherry expression that colocalizes with the microglial marker Iba1. In addition, we utilized flow cytometry to demonstrate time- and Cre-dependent differences in recombination of Cx3cr1+ cells in the spleen, peripheral blood, and brain at two- and eight-weeks post-tamoxifen treatment. Overall, we have created a novel transgenic rat model for researchers to employ in understanding microglial and peripheral immune cell function in rats.

The online version contains supplementary material available at 10.1038/s41598-025-31077-z.