Is this technique published yet?
A manuscript describing the universal insertion sites and miniMos transgenesis has been accepted at Nature Methods (Jan 2014) and should be published around April 2014.
Can I use the reagents prior to publication?
Yes, we have deposited a set of verified universal insertion strains at the CGC. Please see strains and request from the CGC.
Do you use a different protocol to make insertions into the universal insertion sites compared to regular mosSCI?
No, we use exactly the same protocol as we use for “normal” mosSCI insertions. The only difference is that you use the same targeting vector for all the insertion sites and the injection strain determines where the transgene is inserted.
Do I need any new reagents to use the Universal MosSCI sites?
If you have already generated mosSCI insertions into the ttTi5605 site, then you only need to request the new strains for injection. All the co-injection markers are the same. If you have not done any mosSCI experiments, then you will need the reagents described for MosSCI, including a helper plasmid that expresses the mosase (pCFJ601), fluorescent markers to mark the array (pGH8, pCFJ90 and pCFJ104) and a negative selection marker (pMA122). All the reagents have been deposited with Addgene.
What if I want to use the reagents and publish before you get the technique published?
Please cite “Frøkjær-Jensen C, Davis MW, Sarov M, Taylor J, Flibotte S, LaBella M, Pozniakovski A, Moerman DG and Jorgensen EM. Random and targeted transgene insertion in C. elegans using a modified Mos1 transposon. Nature Methods. (In press)”.
Do you have more sites than the standard universal insertion sites?
Yes, we generated a larger set of insertion sites that have not been tested and verified in the same detail as the standard sites. Please contact us (Feedback) if you have a specific use for a site not at the CGC.
Can I make my own universal insertion sites?
Send me an email (see Feedback form) and we can discuss the different options. You can make targeted universal insertion sites by insertion into a Mos1 site, you can generate a random set with the miniMos element or you could make targeted insertion sites by CRISPR.