I am attempting to create multi-copy transgenic arrays of a gene of interest in C. elegans with a fluorescent marker. However I don’t want a bulky GFP tag to interfere with protein function for the gene of interest.
In reading the recent review paper by Jeremy Nance and Christian Frokjaer-Jensen (“The C. elegans Transgenic Toolbox”) they mention an SL1 + SL2 system. However the two citations don’t seem to show an implementation of what I’m thinking of; instead they are general discussions of trans-splicing or nematode genomics. (They cite Cutter et al 2009, and Blumenthal’s 2009 WormBook chapter on operons). While both of these are valuable resources I am still interested in finding out if someone has used, and published the type of operon-based transgenic system I am imagining: specifically,
ubiquitous promoter (plet-858 perhaps)–SL1 [Gene of Interest] --[SL2 GFP] --UTR (unc54? seems like the standard).
Does anyone 1) have something like this or 2) know someone who does? My email address is email@example.com and I would be happy to hear from you! I am aware of some alternative methods for what I’m proposing including 2A self-cleaving peptides. Happy to hear about them as well and whether they work or not. Sounds like it may have some challenges based on the Nance review.