Hello,
I’m trying to do a few experiments with lin-12 and I’d like to test a loss and a gain-of-function allele. Can anyone recommend strains with tips on how they are maintained?
Thank you
I no longer remember the details, but Iva Greenwald’s suppressor/enhancer of lin-12 papers are really good, and you can easily get a sense for what alleles are available, how you can test for modification of lin-12 phenotypes, and which are the strongest viable lin-12 alleles if you want to look for lin-12 phenotypes in some other assay (in addition to the actual null mutant, which is sterile). And as to how to maintain a null allele: it’s available in a balanced strain, and once you replace that balancer with a newer, fluorescently labeled version it will be even easier to maintain.
Hi Kchoe,
Could you tell us more about the experiments you would like to perform ? It could be easier to find the allele that could fit with what you want.
The lin-12 allele can be more or less hard to handle and work with if you are not that familiar with them.
There are many gain and loss of function allele available:
If I remember well one of the strongest gain of function is the n137 allele, but it is really a pain to maintain and work with…
Then you have the n950 allele which is also a really strong gain of function, but it is a bit easier to handle.
As loss of function, I was using a temperature sensitive allele called (n676n930), at 25°C this strain has a pretty good loss of function phenotype.
Hope this helps a bit
This is very helpful; thank you. We have a strong GFP expression phenotype for loss of a gene with homology to DOS proteins and we’d like to test if the mechanism is working through a Notch receptor. We can detect the phenotype by GFP reporter or qPCR. We’d like to see if Notch receptor lf mimics and if a gf suppresses. Is n137 nearly sterile. Is there any temperature sensitivity with either n137 or n950?
I did not work a lot with n137, but with n950.
I haven’t seen any stricking differences in my phenotypes at different temperature but I did not test that in detail.
If you wish to put your marker in the n950 background, I can give you some tips (because as such n950 cannot be crossed), or you can simply reinject your transgene but then your comparison with a WT control will be hard to interpret, you will ork with different arrays.
Do you mean that n950 can’t mate? Thanks, Keith
Indeed, the worms are muv and do not have any functional vulva. And to make the story even more easy, this phenotype is 100% penetrant…
With lin-12(n950gf) I once heat shocked for males, then crossed to another strain to get my double. Not so tough.
In these animals the AC fails to develop, so no vulva = vulvaless. But secondaries are also strongly induced, so there are a lot of small blips. With experience you’ll see that these small little blips are distinguishable from the big blips you get from ectopic primaries. In my strain collection I note n950 and other lin-12(gf) strains as Vul/Mini-Muv.