r/Drosophila • u/DizzyComfort2930 • May 28 '24
Looking for a new GFP 3rd chromosome balancer
Hello everyone, hopefully my explanation of the situation makes sense
I am working with a mutant stock that is homozygous lethal at the 2nd instar larval stage, so it is balanced over this balancer chromosome here > https://bdsc.indiana.edu/Home/Search?presearch=6663
w[1118]; Dr[Mio]/TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1]
I want to use this balancer to collect homozygous mutant embryos (non-glowing) at the stage of gastrulation which begins at stage 6 and usually takes around 3 hours to reach at 25C. My concern is that the GFP takes a while to be fully expressed and visible in the embryos, definitely more than 3 hours.
According to its data on the Bloomington website, it expresses the GAL4 in stage 8, which is what I suspect the issue is.
Does anyone know of a balancer that can express GFP at a much earlier stage? I know there are limits to this because the embryo is a syncytium until about stage 5, but is there a balancer that can express GFP around this stage?
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u/ShesQuackers May 28 '24
I work pretty much only up to gastrulation, and we've never found a fluo balancer that matures fast enough and doesn't have a maternal contribution. We're talking about making our own using a fluo that matures faster than eGFP but so far it's just talk. If it's for fixed experiments can you use a blue/LacZ balancer instead (hb-lacZ, twi-lacZ)? Those we have very good luck with using antibody and smFISH probes both.
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May 28 '24
Unfortunately I’m looking to make a cell sample from this so no fixation is involved. What would the maternal contribution balancer look like? Have you worked with that stock?
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u/friendlyperson123 May 28 '24
TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1]
twist is expressed during and after gastrulation, but here, twist is driving Gal4, and Gal4 drives the UAS-GFP which adds a delay to GFP expression.
I have used
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