ISO panneuronal mCherry (or CFP) insertion with no developmental defects!

Model Organism: Worms Methods & Reagents
1

Hi all,
I’m looking for something rather specific, I suppose but I thought I would give this a try. I’m looking for a transgenic strain that expresses mCherry (or CFP) in most neurons. Panneuronal would be great. So would an unc-86p fusion (or similar). I’d also prefer an insertion although it doesn’t matter where the transgene is inserted (beggars can’t be choosers!). I’ve looked at the CGC and it looks like all the panneuronal mCherry or CFP lines are designed to perturb development in some way. I’d prefer a line that is designed simply to look at neuron morphology.

Of course, I could make this line myself but I’m at a primarily undergraduate institution and so I like to keep my injections/integrations to a minimum. I don’t have time (heavy teaching load)! My current students are not very skilled in this area. Any help would be much appreciated! Thanks.

Maria Gallegos (Cal State University, East Bay)

2

A previous Wormbase Forum thread identifies the following:
gaIs234 [jkk-1p::HIS-24::mCherry + unc-119(+)]
rmIs190 [F25B3.3p::Q67::CFP]
rmIs172 [F25B3.3p::Q19::CFP]
rmIs110 [F25B3.3p::Q40::YFP]
uIs59 [unc-119p::YFP]

Glancing at a couple of obvious genes found more YFP transgenes (not CFP or mCherry, but at least they’re not GFP):
opIs130
irIs37
iwIs25

(By the way, I assume you mean unc-76 and not unc-86)

3

Actually, I did mean unc-86.

Thanks for taking the time to look for these but I forgot to clarify that I do not want nuclear mCherry. Also, the others that you listed I thought would be great but upon closer inspection they have long strings of Qs in them and thus cause neurodegeneration. Sigh.

I’m not sure it’s possible to separate YFP and GFP by microscopy?

Thanks again, though.

4

Most of the newer confocals can separate YFP and GFP via spectral separation (aka linear unmixing). You need to train the microscope using a YFP only and a GFP only strain, and once trained it can recognize the two spectra and thus ‘unmix’ them.

Steve

5

A couple of years ago, I was also looking for a red neuronal line and Morris Maduro kindly sent me the strain MS1180 irIs83[integrant of pMM824 (unc-119::mCherry) and pMM768 (end-3(+)]. I use it for teaching - it’s nice and bright. I would try emailing Morris to see if he can send it to you.

Janet

6

Thanks so much Janet. That sounds promising!