A sequence cassette has been introduced into the attP site present in TI{TI}CCHa1attP, restoring the deleted CCHa1 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 CCHa1 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).
Activation of CCHa1-positive neurons via expression of Bhal\NaChBacUAS.cNa under the control of Scer\GAL4CCHa1-2A-GAL4 leads to a decrease in night time but not day time sleep duration; inhibition via expression of Hsap\KCNJ2UAS.EGFP under the control of Scer\GAL4CCHa1-2A-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.