A sequence cassette has been introduced into the attP site present in TI{TI}TkattP, restoring the deleted Tk sequence and tagging it with 2xTag:MYC. The tag is followed by a 2A-GAL4 cassette, resulting in the GAL4 driver being expressed as a separate protein under the control of the endogenous Tk regulatory sequences. A single loxP site is present downstream of the driver sequence (markers present in the progenitor insertion and donor plasmid have been removed using P1cre).
ScerGAL4Tk-2A-GAL4 drives expression Tachykinin neurons in the adult brain, including in the central complex, the antennal lobes, and the optic lobes, as well as in the ventral nerve cord.
Activation of Tk-positive neurons via expression of Bhal\NaChBacUAS.cNa under the control of Scer\GAL4Tk-2A-GAL4 leads to a significant decrease in day time but not night time sleep duration; inhibition via expression of Hsap\KCNJ2UAS.EGFP under the control of Scer\GAL4Tk-2A-GAL4 leads to a delay in night time but not day time with no changes in sleep duration in day or night time when compared to control adults.
