genotyping tie2:cre (tek-cre#2352rwng) · 2014-10-23 · genotyping tie2:cre (tek-cre#2352rwng) jdw...
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Genotyping Tie2:Cre (Tek-cre#2352Rwng) JDW 3/12 MGI# 3810530 Braren R et al., "Endothelial FAK is essential for vascular network stability, cell survival, and lamellipodial formation." J Cell Biol 2006 Jan 2;172(1):151-62 References: These specific mice are from Rong Wang at UCSF, but the vector she used is the exact same as that originally employed by Tom Sato with the Tie2-promoter driving lacZ followed by the Tie2 enhancer in: Schlaeger, T.M., S. Bartunkova, J.A. Lawitts, G. Teichmann, W. Risau, U. Deutsch, and T.N. Sato. 1997. Uniform vascular-endothelial-cell-specific gene expression in both embryonic and adult transgenic mice. Proc. Natl. Acad. Sci. USA. 94:3058–3063. And the Cre mouse in: Endothelium-specific replacement of the connexin43 coding region by a lacZ reporter gene. Martin Theis1, Cor de Wit2, Thorsten M. Schlaeger3,†, Dominik Eckardt1, Olaf Krüger1, Britta Döring1, Werner Risau3,‡,Urban Deutsch3, Ulrich Pohl2, Klaus Willecke1,*. Genesis. 2001. HHFW3 (5’-GCC TAT GAG AGG ATA CCC CTA TTG) CreRV3 (5’-TCT CTA CAC CTG CGG TGC TAA C) generating a 1kb amplicon spanning the promoter-cre junction. They used the HH-nlscre-XK fragment to make the mice. This is the first use of this as a Cre. Endothelial FAK is essential for vascular network stability, cell survival, and lamellipodial formation. Rickmer Braren 1 , Huiqing Hu 1 , Yun Hae Kim 1 , Hilary E. Beggs 2 , 3 ,Louis F. Reichardt 2 , and Rong Wang 1. 2006. JCB.
JDW 26 (Tie2::Cre-FWD): 5’-CCC TGT GCT CAG ACA GAA ATG AGA (Tie2 FOR) JDW 27 (Tie2::Cre-REV): 5’-CGC ATA ACC AGT GAA ACA GCA TTG C (CRE REV) The transgene produces a 560 bp band. 10X CL Buffer 2.5 95-5:00 Q Solution 2.5 95-0:45 10mM dNTPs 0.5 60-0:30 FWD @ 20 uM 0.5 72-0:30, 35 cycles total REV @ 20 uM 0.5 72-5:00 DNA 1.0 16-forever dH2O 17.25 Q Taq 0.25 *Note: these mice can have widespread Cre activity. It is rare, but using R26RlacZ and mTmG we have seen germline-esque recombination using this allele at a low, but persistent frequency. We highly recommend using a reporter allele in the background if you use this transgene.