B6.Cg-Tg(Thy1-PA-GFP)5Rmpl/J

Stock number: 021069
Research areas: Neurobiology Research, Research Tools

Description

These transgenic mice express nuclear-localized, photoactivatable GFP in neurons, and may be useful to rapidly mark and follow selected populations of neurons in living tissue with single cell resolution using two-photon excitation.

Detailed description

This transgenic strain contains the thymus cell antigen 1 (Thy1) promoter driving expression of a photoactivatable green fluorescent protein, (PA-GFP), fused to a nuclear localization signal (NLS), in neuronal populations. PA-GFP::NLS is enriched in the nucleus, but can be also detected in the cytoplasm of neuronal cells. PA-GFP shows very weak fluorescence at 517nm when excited at 488nm before photoactivation. Upon photoactivation with light at ~400nm PA-GFP is converted into its fluorescent form. This form shows strong emission at 517nm when excited at 488nm and allows the conditional fluorescence labeling of individual, living cells which can be imaged using conventional GFP filter sets. Photoactivation can also be efficiently achieved using 2-photon excitation at ~750nm and imaged after photoconversion using excitation at ~950nm. In these Thy1.2#5 mice, from founder line 5, fluorescence is evident in cortical layer 5 and in fewer neurons in layers 2/3. PA-GFP labeling intensity of individual neurons tends to be strong in these Thy1.2#5 mice as compared to mice from founder line 6 (Stock No. 021070), which shows more even distribution across cortical layers.

Development

This transgene contains a photoactivatable green fluorescent protein, (PA-GFP) fused to a nuclear localization signal (NLS), under the transcriptional control of the thymus cell antigen 1 (Thy1) promoter. The Thy1 promoter used here is a modified regulatory region of the "murine Thy1.2 gene". This construct was injected into fertilized B6.CBAF1 oocytes and mice from founder line 5 were bred to C57BL/6J mice to establish a colony. Transgenic Thy1.2#5 offspring were backcrossed to C57BL/6J mice for at least 8 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Tg(Thy1-PA-GFP)5Rmpl
Name: transgene insertion 5, Simon Rumpel
MGI accession ID: MGI:5449383
Generation method: Transgenic
Attributes: Reporter
Synonyms: Thy1.2#5; Thy1.2-PA-GFP::NLS
Marker symbol: Tg(Thy1-PA-GFP)5Rmpl
Marker name: transgene insertion 5, Simon Rumpel
Chromosome: UN
Strain of origin: (C57BL/6J x CBA)F1
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: Thy1 promoter drives expression of a photoactivatable green fluorescent protein, (PA-GFP), fused to a nuclear localization signal (NLS), in neuronal populations. PA-GFP::NLS is enriched in the nucleus, but can be also detected in the cytoplasm of neuronal cells.
Molecular note: This transgene contains a photoactivatable green fluorescent protein, (PA-GFP) fused to a nuclear localization signal (NLS), under the transcriptional control of the thymus cell antigen 1 (Thy1) promoter. The Thy1 promoter used here is a modified regulatory region of the "murine Thy1.2 gene". Line 5 was generated.