Stocks with donor constructs for making protein traps

The MiMIC collection, generated by the Gene Disruption Project, is composed of a large set of Minos-based insertions carrying an attP-flanked gene trap cassette. This cassette can be swapped with attB-flanked cassettes by phiC31 integrase-catalyzed Recombinase-Mediated Cassette Exchange (RMCE) with simple genetic crosses which combine an excisible swap "donor construct", the necessary recombinase for donor excision, phiC31 integrase and the MiMIC insertion.

The 'Double Header' (DH) donors listed below can be used to make protein traps or gene traps, dependent on the orientation of the final swap. One orientation carries an EGFP-FlAsH-Strep-TEV-3xFlag (GFSTF) tag that, in the correct phase, can be spliced into mRNA and translated.  The other orientation results in a T2A-GAL4 gene trap (see the Trojan GAL4 page for details on how this kind of trap works).

The schematic below (Figure 2 from Li-Kroeger et al., 2018) illustrates how the process works. For more information, see Li-Kroeger et al., 2018 and the Bellen lab page on MiMIC RMCE technology.

For

schematic of dh swap

 




Related link  RMCE for MiMICs