Has anyone made a worm specific sfGFP?

Model Organism: Worms Methods & Reagents
1

Hi,

I am wondering if anyone has made a worm specific sfGFP? Perhaps by mutating one of the Fire vectors?

Thanks,

Vic

2

Yes, we have engineered a worm-codon-optimized superfolder GFP with three operons into a Gateway donor vector. It is detectably brighter in worms than the standard GFP.

Harold

3

Is this reagent published/available to the community? It sounds like a great tool!

best,

Jordan

4

It’s not published, but we’re happy to make it available. Just shoot me an email (smithhe2@niddk.nih.gov) with your shipping address.

Harold

5

Hi Harold. Sounds like a valuable reagent! Do you mean three exons? Also, did you use Fire synthetic introns for this?

6

Hi Harold,

Are you allowed to check your email at this time? I believe you meant 3 introns, not operons, correct? Any data on the half-life of this isoform? Is it the same stability as eGFP or longer? And have you done any assays on how much earlier one can detect expression using this construct vs eGFP?

7

Hi all,

Thanks for catching the operon/intron error; it’s three of the latter (synthetic, 51-bp). Re: Steve’s questions, we made this reagent recently and have only tested its brightness vs. eGFP. There may be some relevant data in the original sfGFP publication (Pédelacq et al., Nature Biotech 2006), but those are in vitro or in coli assays. We don’t intend to investigate these parameters, but you (or anyone else who is interested) are welcome to give it a go.

Harold

P.S.-Now that the furlough is over, I’ll get the plasmid out to those of you who have requested it.