Figure 2.
Genetic elements of the doxycycline-regulated Nkkc1 suppression system. The construct for Actin-tTS::Nkcc1 tetO/tetO. Nkcc1 consists of 26 exons. The translation initiation site (ATG) is situated at exon 1, and the stop site marks the end of translation at exon 26. The tetracycline operator site (tetO) was inserted just upstream of ATG to form Nkcc1 tetO/tetO. Nkcc1 tetO/tetO homozygotes behave like wild-type mice. The tetracycline trans-silencer (tTS) tethers tetO in the absence of doxycycline (DOX), resulting in the suppression of Nkcc1 expression. In the presence of DOX (green circles), tTS-mediated gene knockdown does not occur. Gray boxes: coding regions in exons 1 and 26.