tetOff driver and tetO reporter strains
We have created a larger set of tetOFF driver lines and tetO reporter lines most of which are described in Nonet, 2024 and are available by request. Access the lists below as downloadable excel files which include information about how the lines were made and the sequence of the alleles.
Notes on drivers
Some promoter driver lines express more broadly than the identical promoter fragment expressed as a direct GFP fusions. In most cases we believe this is because the underlying promoters actually express at low level in the other tissues. However, because the bipartite reporter systems are generally limited by the strength of the reporter, not the amount of driver being expressed (see Nonet, 2020), one typically does not see variation in signal strength between cell types as one would with an endogenous promoter. For example, rab-3 promoter fusions appear to express relatively specifically in the nervous system, but rab-3 tetOFF driver tetO nlsGFP strains also express in the intestine. Analysis of single cell RNAseq data confirms that the rab-3 transcript is found in intestinal tissue at low levels.
Notes on reporters
Not all tetO 7X reporter strain express fusions at the same level. Fusions between an FP and endogenous protein expressed at low levels often express at lower levels than the FP alone. Typically, fusions of a poorly expressed protein with poor codon usage with an N-terminal codon optimized FP express better than same fusion with the order reversed. If a fusion protein is bright and does not properly localize, creating new reporter with a lower number of tetO sites will often aid in obtaining a native localization pattern (but will of course, be less bright).
References
Nonet, M.L. Creation and manipulation of bipartite expression transgenes in C. elegans using phiC31 recombinase. bioRxiv 2024.03.01.583017 [link].