Zinc Finger Consortium - Modular Assembly Kits
(Kit #
1000000005, 1000000010
)
Depositing Labs: Keith Joung, Dan Voytas
Zinc finger nucleases (ZFNs) are synthetic proteins commonly used in genome engineering that contain an custom-designed zinc finger DNA-binding domain fused to the cleavage domain of the FokI endonuclease.
Kit #1000000005 includes 141 zinc finger plasmids encoding different zinc finger modules from the Sangamo, Barbas, and Toolgen module sets, and can be used to efficiently assemble zinc finger arrays via modular assembly. Also included in this kit are 5 plasmids and a strain for the bacterial two-hybrid (B2H) system used to screen zinc finger arrays for activity.
This kit will be shipped as bacterial glycerol stocks, 96-well format, on dry ice. Samples should be frozen at -80°C immediately upon arrival.
Kit #1000000010 contains only the B2H reagents, 5 plasmids and 1 strain, and are referred to as the Modular Assembly Accessory Reagents Kit.
This kit consists of five plasmids and one strain that will be shipped as as bacterial stabs at room temperature. IMPORTANT NOTE: all of the materials in this accessory reagents kit are included in the Zinc Finger Consortium Modular Assembly Kit (# 1000000005) and therefore do not need to be purchased in addition to the Modular Assembly Kit.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Kit | 1000000005 | ZFC Modular Assembly Kit - Zinc finger modules (141 plasmids) & bacterial two-hybrid system (5 plasmids + 1 strain) | 1 | $640 | Add to Cart | |
| Kit | 1000000010 | ZFC Accessory Reagents - Bacterial two-hybrid system (5 plasmids + 1 strain) | 1 | $350 | Add to Cart | |
Original Publication
Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc . Nat Protoc. 2006;1(3):1637-52. PubMed PMID 17406455.Description
Zinc fingers are the most common family of transcription factors in organisms ranging from yeast to humans. Custom zinc finger proteins can be designed to target a specific DNA sequence in a gene of interest in nearly any cell type.
Zinc finger nucleases (ZFNs), which consist of an engineered zinc finger DNA-binding domain fused to the DNA cleavage domain of the FokI enzyme, are powerful tools for performing genome engineering in a variety of organisms. ZFNs can introduce double-stranded breaks in DNA that stimulate both homologous and non-homologous recombination pathways to alter a desired locus in the genome.
The zinc finger modules in this kit were derived from the Barbas, Sangamo and ToolGen module sets and standardized to permit "mixing-and-matching" of the different modules. A complementary web-based Zinc Finger Targeter program (ZiFiT) was designed to assist in the identification of potential DNA target sites.
In May 2008, the laboratories of Keith Joung, Daniel Voytas, and Toni Cathomen published a large-scale evaluation of the modular assembly method and reported important guidance for researchers interested in using this approach to engineer multi-finger domains. The work is described in:
Unexpected failure rates for modular assembly of engineered zinc fingers. Ramirez CL, Foley JE, Wright DA, Müller-Lerch F, Rahman SH, Cornu TI, Winfrey RJ, Sander JD, Fu F, Townsend JA, Cathomen T, Voytas DF, Joung JK. Nat Methods . 2008 May;5(5):374-5. doi: 10.1038/nmeth0508-374. PubMed PMID 18446154.Kit Documentation
ZFC Expression Vector and Modular Assembly Kit Info 113.7Â KBLinks
In July 2012, a legacy version of the ZiFiT software package (ZiFiT v3.3) was made available to maintain support for modular assembly of zinc fingers using the Modular Assembly Kit developed by the Zinc Finger Consortium (Wright et al., Nature Protocols 1, 1637-1652, 2006). Considerable progress has been made towards engineering custom DNA binding proteins and the Zinc Finger Consortium now strongly recommends utilizing newer alternative methods for engineering DNA binding domains. For more information on these methods please see the current version of ZiFiT.
Resources
A spreadsheet containing information for the 141 zinc finger plasmids is available as a CSV file:
Zinc Finger Plasmid sequences 761.6Â KBHow to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
"The plasmid kit used for engineering ZFNs was a gift from Keith Joung and Dan Voytas (Addgene kit #1000000005 or #1000000010 )"
For your Reference section:
Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc . Nat Protoc. 2006;1(3):1637-52.
PubMed 17406455.
Zinc Finger Consortium Expression Vector and Modular Assembly Kit - #1000000005
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right, as well as lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin | |
| Kanamycin | |
| Chloramphenicol | |
| Tetracycline |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| Plate 1 / A / 1 | pc3XB-ZF1 | Ampicillin |
| Plate 1 / A / 2 | pc3XB-ZF2 | Ampicillin |
| Plate 1 / A / 3 | pc3XB-ZF3 | Ampicillin |
| Plate 1 / A / 4 | pc3XB-ZF4 | Ampicillin |
| Plate 1 / A / 5 | pc3XB-ZF5 | Ampicillin |
| Plate 1 / A / 6 | pc3XB-ZF6 | Ampicillin |
| Plate 1 / A / 7 | pc3XB-ZF7 | Ampicillin |
| Plate 1 / A / 8 | pc3XB-ZF8 | Ampicillin |
| Plate 1 / A / 9 | pc3XB-ZF9 | Ampicillin |
| Plate 1 / A / 10 | pc3XB-ZF10 | Ampicillin |
| Plate 1 / A / 11 | pc3XB-ZF11 | Ampicillin |
| Plate 1 / A / 12 | pc3XB-ZF12 | Ampicillin |
| Plate 1 / B / 1 | pc3XB-ZF13 | Ampicillin |
| Plate 1 / B / 2 | pc3XB-ZF14 | Ampicillin |
| Plate 1 / B / 3 | pc3XB-ZF15 | Ampicillin |
| Plate 1 / B / 4 | pc3XB-ZF16 | Ampicillin |
| Plate 1 / B / 5 | pc3XB-ZF17 | Ampicillin |
| Plate 1 / B / 6 | pc3XB-ZF18 | Ampicillin |
| Plate 1 / B / 7 | pc3XB-ZF19 | Ampicillin |
| Plate 1 / B / 8 | pc3XB-ZF20 | Ampicillin |
| Plate 1 / B / 9 | pc3XB-ZF21 | Ampicillin |
| Plate 1 / B / 10 | pc3XB-ZF22 | Ampicillin |
| Plate 1 / B / 11 | pc3XB-ZF23 | Ampicillin |
| Plate 1 / B / 12 | pc3XB-ZF24 | Ampicillin |
| Plate 1 / C / 1 | pc3XB-ZF25 | Ampicillin |
| Plate 1 / C / 2 | pc3XB-ZF26 | Ampicillin |
| Plate 1 / C / 3 | pc3XB-ZF27 | Ampicillin |
| Plate 1 / C / 4 | pc3XB-ZF28 | Ampicillin |
| Plate 1 / C / 5 | pc3XB-ZF29 | Ampicillin |
| Plate 1 / C / 6 | pc3XB-ZF30 | Ampicillin |
| Plate 1 / C / 7 | pc3XB-ZF31 | Ampicillin |
| Plate 1 / C / 8 | pc3XB-ZF32 | Ampicillin |
| Plate 1 / C / 9 | pc3XB-ZF33 | Ampicillin |
| Plate 1 / C / 10 | pc3XB-ZF34 | Ampicillin |
| Plate 1 / C / 11 | pc3XB-ZF35 | Ampicillin |
| Plate 1 / C / 12 | pc3XB-ZF36 | Ampicillin |
| Plate 1 / D / 1 | pc3XB-ZF37 | Ampicillin |
| Plate 1 / D / 2 | pc3XB-ZF38 | Ampicillin |
| Plate 1 / D / 3 | pc3XB-ZF39 | Ampicillin |
| Plate 1 / D / 4 | pc3XB-ZF40 | Ampicillin |
| Plate 1 / D / 5 | pc3XB-ZF41 | Ampicillin |
| Plate 1 / D / 6 | pc3XB-ZF42 | Ampicillin |
| Plate 1 / D / 7 | pc3XB-ZF43 | Ampicillin |
| Plate 1 / D / 8 | pc3XB-ZF44 | Ampicillin |
| Plate 1 / D / 9 | pc3XB-ZF45 | Ampicillin |
| Plate 1 / D / 10 | pc3XB-ZF46 | Ampicillin |
| Plate 1 / D / 11 | pc3XB-ZF47 | Ampicillin |
| Plate 1 / D / 12 | pc3XB-ZF48 | Ampicillin |
| Plate 1 / E / 1 | pc3XB-ZF49 | Ampicillin |
| Plate 1 / E / 2 | pc3XB-ZF50 | Ampicillin |
| Plate 1 / E / 3 | pc3XB-ZF51 | Ampicillin |
| Plate 1 / E / 4 | pc3XB-ZF52 | Ampicillin |
| Plate 1 / E / 5 | pc3XB-ZF53 | Ampicillin |
| Plate 1 / E / 6 | pc3XB-ZF54 | Ampicillin |
| Plate 1 / E / 7 | pc3XB-ZF55 | Ampicillin |
| Plate 1 / E / 8 | pc3XB-ZF56 | Ampicillin |
| Plate 1 / E / 9 | pc3XB-ZF57 | Ampicillin |
| Plate 1 / E / 10 | pc3XB-ZF58 | Ampicillin |
| Plate 1 / E / 11 | pc3XB-ZF59 | Ampicillin |
| Plate 1 / E / 12 | pc3XB-ZF60 | Ampicillin |
| Plate 1 / F / 1 | pc3XB-ZF61 | Ampicillin |
| Plate 1 / F / 2 | pc3XB-ZF62 | Ampicillin |
| Plate 1 / F / 3 | pc3XB-ZF63 | Ampicillin |
| Plate 1 / F / 4 | pc3XB-ZF64 | Ampicillin |
| Plate 1 / F / 5 | pc3XB-ZF65 | Ampicillin |
| Plate 1 / F / 6 | pc3XB-ZF66 | Ampicillin |
| Plate 1 / F / 7 | pc3XB-ZF67 | Ampicillin |
| Plate 1 / F / 8 | pc3XB-ZF68 | Ampicillin |
| Plate 1 / F / 9 | pc3XB-ZF69 | Ampicillin |
| Plate 1 / F / 10 | pc3XB-ZF70 | Ampicillin |
| Plate 1 / F / 11 | pc3XB-ZF71 | Ampicillin |
| Plate 1 / F / 12 | pc3XB-ZF72 | Ampicillin |
| Plate 1 / G / 1 | pc3XB-ZF73 | Ampicillin |
| Plate 1 / G / 2 | pc3XB-ZF74 | Ampicillin |
| Plate 1 / G / 3 | pc3XB-ZF75 | Ampicillin |
| Plate 1 / G / 4 | pc3XB-ZF76 | Ampicillin |
| Plate 1 / G / 5 | pc3XB-ZF77 | Ampicillin |
| Plate 1 / G / 6 | pc3XB-ZF78 | Ampicillin |
| Plate 1 / G / 7 | pc3XB-ZF79 | Ampicillin |
| Plate 1 / G / 8 | pc3XB-ZF80 | Ampicillin |
| Plate 1 / G / 9 | pc3XB-ZF81 | Ampicillin |
| Plate 1 / G / 10 | pc3XB-ZF82 | Ampicillin |
| Plate 1 / G / 11 | pc3XB-ZF83 | Ampicillin |
| Plate 1 / G / 12 | pc3XB-ZF84 | Ampicillin |
| Plate 1 / H / 1 | pc3XB-ZF85 | Ampicillin |
| Plate 1 / H / 2 | pc3XB-ZF86 | Ampicillin |
| Plate 1 / H / 3 | pc3XB-ZF87 | Ampicillin |
| Plate 1 / H / 4 | pc3XB-ZF88 | Ampicillin |
| Plate 1 / H / 5 | pc3XB-ZF89 | Ampicillin |
| Plate 1 / H / 6 | pc3XB-ZF90 | Ampicillin |
| Plate 1 / H / 7 | pc3XB-ZF91 | Ampicillin |
| Plate 1 / H / 8 | pc3XB-ZF92 | Ampicillin |
| Plate 1 / H / 9 | pc3XB-ZF93 | Ampicillin |
| Plate 1 / H / 10 | pc3XB-ZF94 | Ampicillin |
| Plate 1 / H / 11 | pc3XB-ZF95 | Ampicillin |
| Plate 1 / H / 12 | pc3XB-ZF96 | Ampicillin |
| Plate 2 / A / 1 | pc3XB-ZF97 | Ampicillin |
| Plate 2 / A / 2 | pc3XB-ZF98 | Ampicillin |
| Plate 2 / A / 3 | pc3XB-ZF99 | Ampicillin |
| Plate 2 / A / 4 | pc3XB-ZF100 | Ampicillin |
| Plate 2 / A / 5 | pc3XB-ZF101 | Ampicillin |
| Plate 2 / A / 6 | pc3XB-ZF102 | Ampicillin |
| Plate 2 / A / 7 | pc3XB-ZF103 | Ampicillin |
| Plate 2 / A / 8 | pc3XB-ZF104 | Ampicillin |
| Plate 2 / A / 9 | pc3XB-ZF105 | Ampicillin |
| Plate 2 / A / 10 | pc3XB-ZF106 | Ampicillin |
| Plate 2 / A / 11 | pc3XB-ZF107 | Ampicillin |
| Plate 2 / A / 12 | pc3XB-ZF108 | Ampicillin |
| Plate 2 / B / 1 | pc3XB-ZF109 | Ampicillin |
| Plate 2 / B / 2 | pc3XB-ZF110 | Ampicillin |
| Plate 2 / B / 3 | pc3XB-ZF111 | Ampicillin |
| Plate 2 / B / 4 | pc3XB-ZF112 | Ampicillin |
| Plate 2 / B / 5 | pc3XB-ZF113 | Ampicillin |
| Plate 2 / B / 6 | pc3XB-ZF114 | Ampicillin |
| Plate 2 / B / 7 | pc3XB-ZF115 | Ampicillin |
| Plate 2 / B / 8 | pc3XB-ZF116 | Ampicillin |
| Plate 2 / B / 9 | pc3XB-ZF117 | Ampicillin |
| Plate 2 / B / 10 | pc3XB-ZF118 | Ampicillin |
| Plate 2 / B / 11 | pc3XB-ZF119 | Ampicillin |
| Plate 2 / B / 12 | pc3XB-ZF120 | Ampicillin |
| Plate 2 / C / 1 | pc3XB-ZF121 | Ampicillin |
| Plate 2 / C / 2 | pc3XB-ZF122 | Ampicillin |
| Plate 2 / C / 3 | pc3XB-ZF123 | Ampicillin |
| Plate 2 / C / 4 | pc3XB-ZF124 | Ampicillin |
| Plate 2 / C / 5 | pc3XB-ZF125 | Ampicillin |
| Plate 2 / C / 6 | pc3XB-ZF126 | Ampicillin |
| Plate 2 / C / 7 | pc3XB-ZF127 | Ampicillin |
| Plate 2 / C / 8 | pc3XB-ZF128 | Ampicillin |
| Plate 2 / C / 9 | pc3XB-ZF129 | Ampicillin |
| Plate 2 / C / 10 | pc3XB-ZF130 | Ampicillin |
| Plate 2 / C / 11 | pc3XB-ZF131 | Ampicillin |
| Plate 2 / C / 12 | pc3XB-ZF132 | Ampicillin |
| Plate 2 / D / 1 | pc3XB-ZF133 | Ampicillin |
| Plate 2 / D / 2 | pc3XB-ZF134 | Ampicillin |
| Plate 2 / D / 3 | pc3XB-ZF135 | Ampicillin |
| Plate 2 / D / 4 | pc3XB-ZF136 | Ampicillin |
| Plate 2 / D / 5 | pc3XB-ZF137 | Ampicillin |
| Plate 2 / D / 6 | pc3XB-ZF138 | Ampicillin |
| Plate 2 / D / 7 | pc3XB-ZF139 | Ampicillin |
| Plate 2 / D / 8 | pc3XB-ZF140 | Ampicillin |
| Plate 2 / D / 9 | pc3XB-ZF141 | Ampicillin |
| Plate 2 / F / 1 | pGP-FF | Ampicillin |
| Plate 2 / F / 2 | pGP-FB-orig BA | Ampicillin |
| Plate 2 / F / 3 | pAC-Kan-alphaGal4 | Kanamycin |
| Plate 2 / F / 4 | pBAC-lacZ | Chloramphenicol |
| Plate 2 / F / 5 | pBAC-BA-lacZ | Chloramphenicol |
| Plate 2 / F / 6 | KJBAC1 strain | Tetracycline |
| Plate 2 / F / 7 | pST1374 | Ampicillin |
| Plate 2 / F / 8 | pDW1775 | Ampicillin |
Kit Plate Map - #1000000005
Zinc Finger Consortium Modular Assembly Accessory Reagents Kit - #1000000010
This kit consists of five plasmids and one strain that will be shipped as as bacterial stabs at room temperature:
| ID | Name |
|---|---|
| 13420 | pGP-FB-orig BA |
| 13421 | pAC-Kan-alphaGal4 |
| 13422 | pBAC-lacZ |
| 13423 | pBAC-BA-lacZ |
| 13424 | KJBAC1 strain |
| 13480 | pGP-FF |