A set of stocks that each contain a Trojan-GAL4 gene trap sequence in a coding intron, inserted via CRISPR/Cas9-induced homology directed repair. The inserted cassette contains a 'Trojan GAL4' gene trap element composed of a splice acceptor site followed by the T2A peptide, the GAL4 coding sequence and an SV40 polyadenylation signal. When inserted in a coding intron in the correct orientation, the cassette should result in truncation of the trapped gene product and expression of GAL4 under the control of the regulatory sequences of the trapped gene. Each line contains one of three versions of the cassette (TI{CRIMIC.TG4.0}, TI{CRIMIC.TG4.1} or TI{CRIMIC.TG4.2}), depending on the frame required to generate a gene trap. In addition, the presence of inverted attP sites flanking the inserted DNA allows for the entire cassette to be replaced with DNA from a compatible donor plasmid (where the sequence to be inserted is flanked by inverted attB sites) through recombination-mediated cassette exchange (RMCE) driven by the phiC31:int integrase.
The PCR check of the insertion gave the expected sized product for the left flank, but no product for the right flank.
The homology arms of the donor vector overlapped by 19 bp. It is unknown whether the insertion is precise or if there is a small deletion or duplication. The insertion site coordinate is the Cas9 cut site predicted for the CRISPR guide RNA.
CR00708
