Hey all, we are just getting into worms in our lab (as in we are about to request our first strains) and I just wanted to get sort of an overall feel for how easy worms are to work with.
I am mostly used to Yeast, so genetic manipulation (KO, tagging, promoter change etc) is very easy, and I am thinking that is not so with worms. Is it easy to clone genes? Just take from cDNA for E. coli? (another thing I never had to deal with is introns!). What sorts of techniques are pretty standardized? I know there is a lot of strains available, so that is pretty nice…though I sure can’t figure out the nomenclature yet! Is N2 the wild-type for basically all strains, or are some sets (like the GFP tag) in a different background? Are they pretty easy to propagate?
This is what I see: deletion in dpy-5(don’t know what e907 is) I am pretty sure that the Co5D10.3 refers to the gene (based on a BAC) and the ::GFP means previous gene is fused to GFP (this is from the GFP list) don’t know what sEX10002, rCes, or +pCeh361 means. I am pretty sure that pCeh361 must be some sort of plasmid in there…
Anyways, hope to be on here to learn alot! Excited to start the worms!
It depends on who you ask. I think it’s safe to say that it’s more challenging than yeast or E. coli, but is still quite manageable and routine. WormBook is an excellent resource for the questions that you ask.
Indeed, dpy-5 is the gene name and e907 is the name of the allele. Right, C05D10.3 is the gene name. If you look up the original reference, it seems that it is a transcriptional fusion, where the promoter of C05D10.3 is being used to drive GFP expression. pCeh361 is a plasmid that contains a wild type copy of the dpy-5 gene, and is used in this case as a co-injection marker for rescue of the e907 lesion.
sEx10002 denotes the extrachromosomal array that was generated by injecting the two constructs together (the transcriptional reporter and the dpy-5 rescue co-injection marker).
Welcome to the worm community! I made the same leap from yeast (albeit some years ago). The biggest difference (for me) was the lack of targeted engineering of strains by homologous recombination. The technology for worms is very much a work in progress, so you’re mostly stuck with traditionally isolated alleles, deletion strains produced by the Knockout Consortium, and/or RNAi. Similarly, transgenes are typically extrachromosomal, multicopy, and prone to germline silencing (although there are ways around these limitations).
On the positive side, worms are far more interesting to study under the microscope (with a host of available GFP/YFP/RFP markers), Wormbase and the CGC strain collection are incredible resources, and wormfolk constitute a community in the truest sense of the word.
Plus, you don’t have to dissect tetrads to get progeny…