Addgene - fox1 (a2bp1l)_R (OZ520) Plasmid Data

Plasmid 27195: fox1 (a2bp1l)_R (OZ520)

Gene/insert name:
Zinc finger array targeting fox1 (a2bp1l)
Alt name:
RNA binding protein, fox-1 homolog (C. elegans) 1-like
Alt name:
OZ520
Alt name:
rbfox1l
Insert size:
270
Species:
D. rerio (zebrafish)
GenBank ID:
ENSDARG00000021184
Entrez Gene:
rbfox1l (Fox-1, fox1l, a2bp1l, hm:zeh0082)
Vector backbone:
MG414
(Search Vector Database)
Vector type:
Zebrafish Targeting
Backbone size w/o insert (bp):
5493
Cloning site 5':
XbaI
Site destroyed during cloning:
No
Cloning site 3':
BamHI
Site destroyed during cloning:
No
5' sequencing primer:
OK.61 GGGTAGTACGATGACGGAACCTGTC List of Sequencing Primers
Bacterial resistance(s)
Kanamycin
Growth strain(s)
XL1 Blue
Growth temperature (℃):
37
Growth instructions:
Requires a bacterial strain such as XL1 Blue, that expresses the lacIq allele of lac repressor
High or low copy:
Low Copy
Sequence: View sequences (1)
Principal Investigator:
Keith Joung
Terms and Licenses:
MTA

Comments:

This plasmid encodes a zinc finger array targeting half of a target sequence in the zebrafish gene fox1 (a2bp1l) (see below). Please note that this plasmid does NOT contain the fox1 (a2bp1l) sequence.

Users must order the complementary plasmid fox1 (a2bp1l)_L (OZ519) [Addgene plasmid 27194] 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'-GCTGAGGGC-3' in a bacterial two hybrid assay, and resulted in 8.09 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

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification. Maeder et al (Mol Cell. 2008 Jul 25. 31(2):294-301. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 27195" in your Materials and Methods section.