The following information accompanied stocks donated to the Bloomington Stock Center by Rachel Wilson, Harvard University.
UAS-driven cytosolic mStayGold and membrane-targeted mStayGold. Bright, photostable GFP reagent that is useful for live imaging, particularly during development. When membrane tagged it is superior to mCD8::GFP for brightness, which is great for patch-clamp of 'dim' driver lines.
P{20XUAS-IVS-mStayGold}su(Hw)attP5
PBac{20XUAS-IVS-mStayGold}VK00005
P{20XUAS-IVS- mCD8::mStayGold }su(Hw)attP5
PBac{20XUAS-IVS- mCD8::mStayGold }VK00005
UAS-driven cytosolic mNeonGreen. Reagent is super-resolution compatible and seems to work for live-cell super resolution (e.g. STED microscopy).
P{20XUAS-IVS-mNeonGreen}su(Hw)attP5
PBac{20XUAS-IVS-mNeonGreen}VK00005
mCyRFP3-Kir fusions: Kir2.1 fused to a cyan-excitable red-fluorescent protein (mCyRFP3) that is visible during standard 2-photon imaging without extra hardware. All were validated by patch clamp and functional imaging.
P{20XUAS-IVS- mCyRFP3::Kir2.1 }su(Hw)attP8
PBac{20XUAS-IVS- mCyRFP3::Kir2.1 }VK00002
PBac{20XUAS-IVS- mCyRFP3::Kir2.1 }VK00033
PBac{13XlexAop-IVS- mCyRFP3::Kir2.1 }VK00002
PBac{13XlexAop-IVS- mCyRFP3::Kir2.1 }VK00033
mCyRFP3-CsChrimson fusions: optogenetic stock with CsChrimson fused to mCyRFP3 that is visible during standard 2-photon imaging. All validated by patch clamp/2-photon for light-activation, and wide-field/2-photon for fluorophore localization and brightness.
PBac{20XUAS-IVS- CsChrimson::mCyRFP3 }VK00002
PBac{13XlexAop-IVS- CsChrimson::mCyRFP3 }VK00002
mCyRFP3-GCaMP fusions and bicistronic expression constructs: fused GCaMP7 and GCaMP8 variants with mCyRFP3 for ratiometric imaging. Also created bicistronic constructs for experiments that require the expression of both constructs separately. All have been validated for expression and function, in some cases with a large number of drivers.
PBac{20XUAS-IVS- jGCaMP7f::mCyRFP3 }VK00033
P{20XUAS-IVS-jGCaMP7f-T2A-mCyRFP3}su(Hw)attP5
PBac{20XUAS-IVS-jGCaMP7f-T2A-mCyRFP3}VK00005
PBac{20XUAS-IVS- jGCaMP8s::mCyRFP3 }VK00033
PBac{20XUAS-IVS-jGCaMP8s-T2A-mCyRFP3}VK00005
PBac{20XUAS-IVS- jGCaMP8m::mCyRFP3 }VK00033
P{20XUAS-IVS-jGCaMP8m-T2A-mCyRFP3}su(Hw)attP5
PBac{20XUAS-IVS-jGCaMP8m-T2A-mCyRFP3}VK00005
Syt1-mCyRFP3-GCaMP double fusions and bicistronic expression constructs: synapse-targeted calcium indicator with mCyRFP3 fluorophore. Also created bicistronic constructs for experiments that require the expression of both constructs separately. These have been validated by functional imaging.
PBac{20XUAS-IVS- Syt1::jGCaMP8m::mCyRFP3 }VK00033
PBac{20XUAS-IVS- Syt1::jGCaMP8m-T2A-Syt1::mCyRFP3 }VK00005
Further details on the generation and validation of these lines is available in the Associated file for this personal communication.