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).

MiMIC insertions inside coding introns can be used to make protein traps by swapping in an artificial exon carrying GFP, RFP or other protein tags. Artificial exons in the right orientation and reading frame (or "phase") can be spliced into mRNA and translated, resulting in a tagged protein.

Swaps can be now be accomplished with simple genetic crosses which combine an excisible swap "donor construct", the necessary recombinase for donor excision, phiC31 integrase and the MiMIC insertion. The schematic below illustrates how the process works using a FRT-excisible donor carrying an EGFP-FlAsH-Strep-TEV-3xFlag (GFSTF) tag generated by the Bellen lab (Nagarkar-Jaiswal et al., 2015). For more information and crossing schemes see Nagarkar-Jaiswal et al., 2015 and the Bellen lab page on MiMIC RMCE technology.

DNA carrying a GFSTF exon is excised using FLP-mediated recombination and then swapped into the MiMIC insertion using phiC31-mediated recombination.

Below is a list of donor constructs for swapping. For a list of already-generated MiMIC RMCE protein traps see here.



Related link  RMCE for MiMICs