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Species: Danio rerio
Genetic Insert: Zinc finger array targeting morc3b
Vector Backbone Description: Backbone Size:5493; Vector Backbone:MG414; Vector Types:Zebrafish Targeting; Bacterial Resistance:Kanamycin
Defining Citation: PMID:18657511
Comments: This plasmid encodes a zinc finger array targeting half of a target sequence in the zebrafish gene morc3b. Please note that this plasmid does NOT contain the morc3b sequence.
Users must order the complementary plasmid morc3b_L (OZ575) [Addgene plasmid 35215] 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), pMLM290/292 (Addgene plasmids 21872 & 21873), pMLM800/802 (Addgene plasmids 27202 & 27203).
The difference between pMLM800/802 and pMLM290/292 is the length of the “spacer” sequence in the full ZFN target site. If the spacer is 7 bp, scientists should use pMLM800/802. If the spacer is 5 or 6 bps, scientists should use pMLM290/292.
pMLM290/292 is identical to pST1374 except that it harbors two mutations in the FokI nuclease domain (Q486E, I499L; aka the “-“ mutation see Miller et al., Nat. Biotech 2007, PMID 17603475) which confers heterodimeric behavior on these domains
This zinc finger array was tested for binding activity to the sequence 5'-GCTGGAGGA-3' in a bacterial two hybrid assay, and resulted in 10.9 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 [email protected] 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)
Proper citation: RRID:Addgene_35216 Copy
Species: Danio rerio
Genetic Insert: Zinc finger array targeting zgc:162148
Vector Backbone Description: Backbone Size:5493; Vector Backbone:MG414; Vector Types:Zebrafish Targeting; Bacterial Resistance:Kanamycin
Defining Citation: PMID:18657511
Comments: This plasmid encodes a zinc finger array targeting half of a target sequence in the zebrafish gene zgc:162148. Please note that this plasmid does NOT contain the zgc:162148 sequence.
Users must order the complementary plasmid zgc:162148_R (OZ580) [Addgene plasmid 35220] 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), pMLM290/292 (Addgene plasmids 21872 & 21873), pMLM800/802 (Addgene plasmids 27202 & 27203).
The difference between pMLM800/802 and pMLM290/292 is the length of the “spacer” sequence in the full ZFN target site. If the spacer is 7 bp, scientists should use pMLM800/802. If the spacer is 5 or 6 bps, scientists should use pMLM290/292.
pMLM290/292 is identical to pST1374 except that it harbors two mutations in the FokI nuclease domain (Q486E, I499L; aka the “-“ mutation see Miller et al., Nat. Biotech 2007, PMID 17603475) which confers heterodimeric behavior on these domains
This zinc finger array was tested for binding activity to the sequence 5'-GCTGCTGGA-3' in a bacterial two hybrid assay, and resulted in 3.1 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 [email protected] 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)
Proper citation: RRID:Addgene_35219 Copy
Species: Danio rerio
Genetic Insert: Glial fibrillary acidic protein promoter
Vector Backbone Description: Backbone Size:3323; Vector Backbone:pminiTol2; Vector Types:Tol2 mediated recombination; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28882119
Proper citation: RRID:Addgene_178963 Copy
Species: Danio rerio
Genetic Insert: Glial fibrillary acidic protein promoter
Vector Backbone Description: Backbone Size:3323; Vector Backbone:pminiTol2; Vector Types:Tol2 mediated recombination; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28882119
Proper citation: RRID:Addgene_178964 Copy
Species: Danio rerio
Genetic Insert: Histone deacetylase 6
Vector Backbone Description: Vector Backbone:pAC8; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:36255643
Proper citation: RRID:Addgene_180226 Copy
Species: Danio rerio
Genetic Insert: TagRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117766 Copy
Species: Danio rerio
Genetic Insert: TagRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-TagBFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117768 Copy
Species: Danio rerio
Genetic Insert: TagRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2329; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Control integrates at the gene cx43.4 using the guide RNA Target "5'-GGGCCATATCTTGCCCACGA-3'" for wide-spread RFP expression in zebrafish embryos from 12hrs post injection through 5 days development
Proper citation: RRID:Addgene_117801 Copy
Species: Danio rerio
Genetic Insert: Cre
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-TagBFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117792 Copy
Species: Danio rerio
Genetic Insert: Cre
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-eGFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117791 Copy
Species: Danio rerio
Genetic Insert: Cre
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117793 Copy
Species: Danio rerio
Genetic Insert: mRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117784 Copy
Species: Danio rerio
Genetic Insert: mRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-TagBFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117786 Copy
Species: Danio rerio
Genetic Insert: gal4/VP16
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117781 Copy
Species: Danio rerio
Genetic Insert: Cre
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-TagBFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117789 Copy
Species: Danio rerio
Genetic Insert: mRFP
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-eGFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117788 Copy
Species: Danio rerio
Genetic Insert: Cre
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117790 Copy
Species: Danio rerio
Genetic Insert: 3x3 Stop sequence
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-eGFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117773 Copy
Species: Danio rerio
Genetic Insert: 3x3 Stop sequence
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a cardiac-myosin-light-chain-2 promoter driving NLS-mRFP, Oline tools and cloning help avalible at
Proper citation: RRID:Addgene_117775 Copy
Species: Danio rerio
Genetic Insert: 3x3 Stop sequence
Vector Backbone Description: Backbone Marker:Karl J. Clark; Backbone Size:2856; Vector Backbone:pK; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32412410
Comments: Secondary gene is a gamma-crystalin promoter driving NLS-mRFP. Online tools and cloning help available at
Proper citation: RRID:Addgene_117772 Copy
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