I'm trying to express transmembranal FasL in cells that do not express it naturally and i'm looking for a plasmid that i can use.
Looked in addgene and it's not there.
In the literature I find only soluble FasL expression but I would like to have it transmembrane bound.
Hi Lior,
when you say "membranal" - do you mean transmembrane, or membrane-associated through a motif to a location vicinal to the cell membrane?
In the latter case, one possibility could be to generate something like a C-terminal prenylation CAAX motif / polybasic motif into a reading frame for FasL as has been done with quite a few subcellular localisation/translocation studies over the years (ie Choy et al 1999, https://www.ncbi.nlm.nih.gov/pubmed/10412982/) with the Ras family - whether this would work for FasL is a different question (it is outwith my field). If it is not available at Addgene or by contacting another lab in the field who have published a similar construct, this kind of fusion protein could easily be generated by gene synthesis and subcloned into a mammalian expression vector for your experiments if you know or can define the sequence(s) you want to work with.
Or - having just quickly read some FasL literature - are you looking for full-length transmembrane Fas Ligand CD95L? If a construct isn't available direct from the several groups who have worked on this, rather than Addgene, you would still be able to get this made by gene synthesis.
Good luck with your project!
Thank you Robin Dickinson for your answer.
I am looking to express to full length transmembrane Fas Ligand (CD95L).
Going through a lot of literature, I didn't find a lab to ask it from and gene synthesis is indeed a viable solution.
Still hope to find a pre-made plasmid.
Thank you!
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