Tg(UBC-Xrcc4,-GFP)1Jpdv

This transgene was produced by transfection of single-cell embryos by a self-inactivating lentiviral vector with a truncated 3' LTR into which a loxP site has been introduced; upon integration, the 3' LTR is duplicated at the 5' end of the insertion so that the entire transgene can be excised by Cre recombinase, leaving behind a single loxP-delta-3' LTR. The vector contains the human ubiquitin C promoter upstream of the mouse Xrcc4 (X-ray repair complementing defective repair in Chinese hamster cells 4) open reading frame joined, in-frame, to a DNA fragment encoding a tandem affinity purification (TAP) tag -- which consists of a calmodulin binding peptide joined via a linker sequence containing a TEV protease cleavage site to two copies of the protein A IgG binding domain -- followed by an internal ribosomal entry site (IRES2) and the coding sequence for green fluorescent protein. Southern blot analysis demonstrated the presence of a single proviral integration, which was mapped by inverse PCR analysis to Chr 16, in the third intron of the Spag6 (sperm associated antigen 6) gene. Expression of the XRCC4 protein in spleen and thymus was demonstrated by immunoblot analysis, and the transgene's complementation of the embryonic lethal brain neuronal apoptosis phenotype of Xrcc4tm1Alt homozygous mice demonstrates XRCC4 expression in the brain; GFP is not expressed in mice with this transgene. (J:125880, J:181755)