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URL: http://www.addgene.org/40591

Proper Citation: RRID:Addgene_40591

Insert Name: Cre recombinase

Organism: Homo sapiens

Bacterial Resistance: Ampicillin

Defining Citation: PMID:11668683

Vector Backbone Description: Backbone Marker:Michael Brenner (PMID: 7952266); Backbone Size:5500; Vector Backbone:pgfa2-lac2; Vector Types:Mammalian Expression, Mouse Targeting, Cre/Lox; Bacterial Resistance:Ampicillin

Comments: This plasmid encodes a transgene to express the Cre recombinase in astrocytes of transgenic mice. A DNA fragment encoding the Cre recombinase was inserted into an expression cassette containing the gfa2 promoter (Besnard et al., 1991), a 2.2-kb 5' flanking region from the human GFAP (hGFAP) gene. The nuclear targeting signal from the SV40 large T antigen was inserted at the amino terminal end of the coding region, because it reportedly increases the efficiency of DNA excision by the recombinase (Gu et al., 1993). A heterologous intron and polyadenylation signal were provided by sequences from the mouse protamine 1 gene at the 3' end. The hGFAP-cre transgene was constructed by excising the lacZ coding region from pgfa2-lac2 (Brenner et al., 1994) by BamHI digestion, and replacing it by blunt end ligation with a nuclear-targeted Cre coding sequence. The latter was obtained as a 1.1-kb SalI-MluI fragment from plasmid pTZ19RCreNLS, which was kindly provided by Drs. Raul Torres and Klaus Rajewsky. Both junctions were verified by DNA sequencing.

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