( B) Representative images of native EGFP and tdTomato fluorescence in VISp of Cre Ai139 double transgenic mice. The tTA2 activity and subsequent TRE2 promoter-driven expression can be suppressed by giving doxycycline (DOX) to double transgenic mice. ( A) Cre-dependent TIGRE2.0 reporter line design and simplified double transgenic breeding scheme. The extent of reporter expression in inhibitory neurons varied from none (Plxnd1-IRES2-dgFlpO, Rorb-P2A-FlpO), to a few cells (Rasgrf2-T2A-dgFlpO), to many cells (Calb1-IRES2-Cre). Representative images of dFISH in VISp, using tdTomato reporter (green) and pan-GABAergic marker Gad1 (red) probes, are shown. ( B) Examination of excitatory and inhibitory neuronal labeling in example cortical layer-specific driver lines. The only exception is Tnnt1-IRES2-CreERT2, which was crossed to a new TIGRE2.0 reporter Ai140 and TissueCyte imaging shows extremely sparse GFP labeling of thalamic neurons (after 1-day tamoxifen induction at P10). Ai14 was used to examine all Cre lines, and Ai65F was used for all FlpO lines. ( A) Selected images from ISH characterization of all new driver lines using either tdTomato reporter probe or Cre probe as indicated on each panel. These novel transgenic lines greatly expand the repertoire of high-precision genetic tools available to effectively identify, monitor, and manipulate distinct cell types in the mouse brain.Ĭre Flp TIGRE calcium sensor cell type channelrhodopsin optogenetics reporter transgenic mice voltage sensor.Ĭopyright © 2018 Elsevier Inc.

TIGRE2.0 reporters broke the barrier in transgene expression level of single-copy targeted-insertion transgenesis in a wide range of neuronal types, along with additional advantage of a simplified breeding strategy compared to our first-generation TIGRE lines.

In particular, we describe the TIGRE2.0 transgenic platform and introduce Cre-dependent reporter lines that enable optical physiology, optogenetics, and sparse labeling of genetically defined cell populations. Here, we report a large set of driver and reporter transgenic mouse lines, including 23 new driver lines targeting a variety of cortical and subcortical cell populations and 26 new reporter lines expressing an array of molecular tools. Modern genetic approaches are powerful in providing access to diverse cell types in the brain and facilitating the study of their function.

Mouse driver