The JFRC UAS transgene goes v3.0
I've blogged a bit on the very useful upgrades to the Gal4/UAS system that has come out of JFRC so far. I think the latest round of tweaks makes the UAS construct so much better that it could be called the 3.0 version.
Earlier, Pfeiffer et al. introduced a UAS construct that most notably contained the IVS and WPRE elements as upgrades to the bland looking 1.0 version (in retrospect). I actually jumped on the v2.0 wagon (which reads 10xUAS-IVS-GFP-WPRE-SV40term.) and quickly made UAS-dWGA:GFP and UAS-dPNA:GFP fly stocks towards developing a good trans-synaptic marker. I must say it paid off nicely! Well, just when I was thinking "OK, this should last a while", it got even better!
Upon RTFA it was easy to see why they have been trying to push the Gal4/UAS based transgene expression amplitude even higher. May be it also has to do with the weak driving capacity of many DSCP-Gal4 lines? I wonder... Anyway, I also applaud at the effort to overcome the endogenous level of Shibire activity with the robust level of transgenic UAS-Shibire(ts1) expression through the introduction of some really cool design changes. We noted this requirement of high Shibire(ts1) expression level as well recently for silencing larval sensory and motor neurons (bummer.. missed out on a cite).
In summary, if you want to express a cDNA of interest at an epic level using the Gal4/UAS system, then 10xUAS-IVS-syn21-cDNA-p10 is the way to go. For this, one could simply ligate a [RE1]-syn21-seq-AATCAAA-cDNA-[RE2] PCR product into a vector backbone that reads: 10xUAS-IVS-MCS-p10. I think such a backbone will be very useful to make.. may be it will also be useful to flank it with the Gypsy insulator elements to keep it extra quiet when there is no Gal4 around. I'd then call it v3.5! Finally, I must thank Barret for the e-mail discussions.
Direct link to Pfeiffer et al. 2012.
PS: This blog is now being mirrored at balapagos.wordpress.com. This is in case Twitter decides to gradually decommission Posterous, of which there have been some indications recently.






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