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Table 1 Viral combinations used in this study and their functions.

From: Cell- and region-specific miR30-based gene knock-down with temporal control in the rat brain

Abbreviation Vector combination Function
LVVs-miRLuc-neurone LV-SYN-Luc
+ LV-Tretight-GFP-miR30-shRNA/Luc
+ LV-mCMV/SYN-tTA
Neurone-specific Luc knock-down system. Tet-off transactivator tTA is expressed from LV-mCMV/SYN-tTA which contains the bidirectional amplified SYN promoter to ensure high level of tTA. tTA binds to Tretight promoter in LV-Tretight-GFP-miR30-shRNA/Luc and activates the expression of shRNA/Luc.
LVVs-miRLuc-glia LV-GfaABC1D-Luc
+LV-Tretight-GFP-miR30-shRNA/Luc
+LV-mCMV/GfaABC1D-tTA
Astrocyte-specific Luc knock-down system. Transactivator tTA is expressed from LV-mCMV/GfaABC1D-tTA which contains the bidirectional amplified GfaABC1 D promoter to ensure high level of tTA. tTA binds to Tretight promoter in LV-Tretight-GFP-miR30-shRNA/Luc and activates the expression of shRNA/Luc.
LVVs-miRLuc-control1 LV-SYN-Luc
+ LV-Tretight-GFP-miR30-shRNA/Luc
+ LV-SYN-WPRE
Control combination used in Luc knock-down experiments for the neurone-specific system. Since there is no tTA expression in this combination, shRNA/Luc will not be expressed. So this combination marked the background expression level of Luc driven by SYN promoter.
LVVs-miRLuc-control2 LV-GfaABC1D-Luc
+ LV-Tretight-GFP-miR30-shRNA/Luc + LV-GfaABC1D-WPRE
Control combination used in Luc knock-down experiments for the astrocyte-specific system. Similar as LVVs-miRLuc-control 1, there is no tTA expression in this combination; shRNA/Luc will not be expressed. Therefore this combination marked the background expression level of Luc driven by GfaABC1 D promoter.
LVVs-miRnNOS-neurone LV-Tretight-GFP-miR30-shRNA/nNOS
+ LV-mCMV/SYN-tTA
Neurone-specific nNOS knock-down system. Mechanism is similar as LVVs-miRLuc-neurone.
LVVs-miRnNOS-glia LV-Tretight-GFP-miR30-shRNA/nNOS + LV-mCMV/GfaABC1D-tTA Astrocyte-specific Luc knock-down system. Mechanism is similar as LVVs-miRLuc-glia.
LVVs-miRnNOS-control1 LV-Tretight-GFP-miR30-shRNA/nNOS + LV-SYN-WPRE Control combination used in nNOS knockdown experiments for the neurone-specific system. Mechanism is similar as LVVs-miRLuc-control1.
LVVs-miRnNOS-control2 LV-Tretight-GFP-miR30-shRNA/nNOS + LV-GfaABC1D-WPRE Control combination used in nNOS knockdown experiments for the astrocyte-specific system. Mechanism is similar as LVVs-miRLuc-control2.
LVVs-miRnNOS-negative control1 LV-Tretight-GFP-miR30-shRNA/Luc
+ LV-mCMV/SYN-tTA
Negative control combination used in nNOS knock-down experiments for the neurone-specific system. Instead of producing shRNA/nNOS it produces shRNA/Luc which shouldn't work on nNOS knockdown.
LVVs-miRnNOS-negative control2 LV-Tretight-GFP-miR30-shRNA/Luc
+ LV-mCMV/GfaABC1D-tTA
Negative control combination used in nNOS knock-down experiments for the astrocyte-specific system. Instead of producing shRNA/nNOS it produces shRNA/Luc which shouldn't work on nNOS knockdown.