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Koretzky Laboratory

Summary:

Our laboratory makes use of cell-free biochemical systems, model cell lines, and whole animals which have been genetically manipulated to probe various signal transduction pathways. Over the past several years we have become particularly interested in the regulation and integration of second messenger cascades by adapter molecules, those proteins which possess no intrinsic enzymatic properties, but which function by bridging protein-protein interactions.

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Organisms and Viruses

  • Akt2fl/fl ( Mus musculus )

    "An frt-flanked PGK-neomycin resistance cassette was inserted into intron 5, and loxP sites were inserted flanking exons 4 and 5."

  • B6.Cg-Akt2(tm1.1Mbb/J) ( Mus musculus )

    "Mice were generated with a targeted disruption in the Akt2 locus by homologous recombination. The targeting vector was designed to insert LoxP sites flanking the sequence containing the coding exons 4 and 5 Mice harboring the targeted allele were identified by Southern blotting and were mated to transgenic mice expressing Cre recombinase.
    The progeny carrying both the Cre transgene and the targeted allele were mated with wild-type (WT) mice to obtain offspring in which the Cre transgene was segregated away and the targeted allele was excised, as determined by the polymerase chain reaction (PCR) and Southern blotting, respectively. These mice were mated inter se to produce offspring with homozygous deletions of Akt2."

  • DGKzeta-deficient mice ( Mus musculus )

    " DGKzeta-deficient mice were generated using homologous recombination in mouse embryonic stem cells. In the targeted Dgkz allele, a 6.1-kb genomic DNA fragment, encompassing exons encoding the C1 and kinase domains, was replaced by the neomycin resistance gene and a cassette of internal ribosomal entry site and green fluorescent protein (IRES-GFP) to mark cells normally expressing DGKzeta."

    Mice are backcrossed 10 generation to C57Bl/6.

  • Hif1fl/fl ( Mus musculus )

    "A loxP site was inserted in intron 1 and a floxed neomycin resistance cassette was placed in intron 2. Exon 2 was left flanked by loxP sites after the neo cassette was excised by in vitro expression of cre recombinase."

  • MTS knockin mice ( Mus musculus )

    "Base pairs 1–105 of murine LAT cDNA were fused to LCP2 exon 1 and intron 1 DNA sequences. The LCP2 ATG was mutated, and the resulting fragment was cloned into the pPNT-FRT vector adjacent to the FRT-flanked neoR cassette. The targeting construct was electroporated into R1 embryonic stem cells, and injected into B6 × 129 blastocysts. The neoR cassette was excised by breeding to FLPe mice."

  • SLP-76+/− ( Mus musculus )

    "About 360 base pairs (bp) of the SLP-76 genomic locus, including 145 bp of the first exon containing the translational start site, were replaced with a neomycin resistance cassette in the reverse transcriptional orientation.
    Two of the clones were microinjected into C57BL/6 blastocysts, and chimeric mice were then bred to wild-type C57BL/6 mice. "

  • SLP-76F/FCD4cre ( Mus musculus )

    "SLP-76F/F mice were crossed with CD4cre transgenic mice to generate SLP-76F/FCD4cre mice."

  • SLP76-KO ( Mus musculus )

    "About 360 base pairs (bp) of the SLP-76 genomic locus, including 145 bp of the first exon containing the translational start site, were replaced with a neomycin resistance cassette in the reverse transcriptional orientation.
    Two of the clones were microinjected into C57BL/6 blastocysts, and chimeric mice were then bred to wild-type C57BL/6 mice. Heterozygous mice were then mated to obtain homozygous SLP-76–deficient mice."

  • Y112/128F ( Mus musculus )

    Mice were generated expressing SLP76 containing a double mutation in Y112/128 to phenylalanine.

  • Y145F ( Mus musculus )

    Mice were generated expressing SLP76 containing a single mutation in Y145 to phenylalanine (Y145F mice).

  • “floxed” SLP-76 mice ( Mus musculus )

    "Mice were generated in which exon 3 of the SLP-76 gene was flanked by loxP sites (floxed)".


Last updated: 2015-09-28T09:42:12.120-04:00

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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016