The endogenous TBPH coding region is replaced with fly codon-optimized Hsap\TARDBP cDNA with an N-terminal Tag:FLAG tag and a Disc\RFPDsRed.3xP3.cUa marker flanked by loxP sites at the C-terminal. This results in the Hsap\TARDBP sequence being expressed under the control of the endogenous TBPH regulatory regions.
TBPHKO.hTARDBP.WT.DsR+ homozygotes of either sex are viable and display no significant changes in developmental timing, longevity or fecundity compared to controls. Neither larval (distance travelled in 5min) not adult locomotion is perturbed (the adults show normal diurnal activity pattern and both the overall daily activity and climbing assay performance as they age is comparable to controls).