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URL: http://www.addgene.org/27189
Proper Citation: RRID:Addgene_27189
Insert Name: Zinc finger array targeting PTRF/Cavin1
Organism: Danio rerio
Bacterial Resistance: Kanamycin
Defining Citation: PMID:18657511
Vector Backbone Description: Backbone Size:5493; Vector Backbone:MG414; Vector Types:Other, Zebrafish Targeting; Bacterial Resistance:Kanamycin
Comments: This plasmid encodes a zinc finger array targeting half of a target sequence in the zebrafish gene PTRF/Cavin1 (see below). Please note that this plasmid does NOT contain the PTRF/Cavin1 sequence. Users must order the complementary plasmid PTRF/Cavin1_L (OZ513) [Addgene plasmid 27188] in order to create a zinc finger nuclease (ZFN) pair to introduce targeted mutations into this specific zebrafish gene. Users will also need to clone the zinc finger (ZF) insert of this plasmid into a zinc finger nuclease (ZFN) vector to express a FOKI fusion product. Examples of possible ZFN expression vectors that can be used are: pST1374 (Addgene plasmid 13426), pMLM290 (Addgene plasmid 21872), pMLM292 (Addgene plasmid 21873), pMLM800 (Addgene plasmid 27202) and pMLM802 (Addgene plasmid 27203). This zinc finger array was tested for binding activity to the sequence 5'-GGAGGTGCC-3' in a bacterial two hybrid assay, and resulted in 6.81 fold activation. However, this array has not yet been tested for activity as a zinc finger nuclease (i.e. for its ability to induce mutations at the intended locus). Scientists using this zinc finger array in a publication should notify zebrafishzfs@zincfingers.org and acknowledge NIH grant number R01 GM088040 in the publication. Other Articles: "Oligomerized pool engineering (OPEN): an 'open-source' protocol for making customized zinc-finger arrays." Maeder ML et al. (Nat Protoc. 2009 Sept 17. 4(10):1471-1501. Pubmed ID: 19798082 "Targeted mutagenesis in zebrafish using customized zinc-finger nucleases." Foley JE et al. (Nat Protoc. 2009 Dec 3. 4(12):1855-1867. Pubmed ID: 20010934
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Source: Addgene
