Wolfe-Lawson ZFN modular assembly kit
(Kit #
1000000018
)
Depositing Labs: Scot Wolfe, Nathan Lawson
This plasmid kit contains reagents for engineering zinc finger arrays by modular assembly method.
This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80°C immediately upon arrival. Individual plasmids can be ordered from each plasmid page and will be shipped as bacterial stabs.
Original Publication
Evaluation and application of modularly assembled zinc finger nucleases in zebrafish. Zhu C, Smith T, McNulty J, Rayla AL, Lakshmanan A, Siekmann AF, Buffardi M, Meng X, Shin J, Padmanabhan A, Cifuentes D, Giraldez AJ, Look AT, Epstein JA, Lawson ND, and Wolfe SA. Development 2011, 138:4555-4564. PubMed
Description
Zinc-finger nucleases (ZFNs) allow targeted gene inactivation in a wide range of model organisms. We provided an archive of 81 clones containing 27 different zinc-finger modules at finger 1, 2 and 3 positions for straightforward construction of ZFNs using modular assembly-based approach. The majority of zinc-finger proteins assembled from these modules have favorable specificities and nearly one-third of modular ZFNs generated lesions at their targets in the zebra fish genome. These modules are designed for the construction of three-finger zinc-finger ZFNs, but they can also be used for assembling longer ZFNs containing 4 or more modules.
Figure : Modules containing in this archive can recognize the following triplet target sites (5’→3’):
Kit Documentation
Wolfe-Lawson ZFN kit 286.1Â KBResources
We set up a website to help users search appropriate sites in their target sequences and design corresponding ZFNs using this archive:
http://pgfe.umassmed.edu/ZFPmodularsearch.html
To facilitate the application of ZFNs within the zebrafish community we constructed a public database of sites in the zebrafish genome that can be targeted using this archive:
Protocols
Protocols for modular assembly construction of ZFNs can be found at the above website as well as in the publication.
How 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 generation of ZFNs was a gift from Scot Wolfe and Nathan Lawson (Addgene kit # 1000000018)"
For your Reference section:
Evaluation and application of modularly assembled zinc finger nucleases in zebrafish. Zhu C, Smith T, McNulty J, Rayla AL, Lakshmanan A, Siekmann AF, Buffardi M, Meng X, Shin J, Padmanabhan A, Cifuentes D, Giraldez AJ, Look AT, Epstein JA, Lawson ND, and Wolfe SA. Development 2011, 138:4555-4564.
PubMed
Wolfe-Lawson ZFN modular assembly kit - #1000000018
- 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 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 |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pBS-F1-AAG | Ampicillin |
| A / 2 | pBS-F1-ACG | Ampicillin |
| A / 3 | pBS-F1-AGG | Ampicillin |
| A / 4 | pBS-F1-ATG | Ampicillin |
| A / 5 | pCS2-F1-CAG-F2-GAC | Ampicillin |
| A / 6 | pBS-F1-CGG | Ampicillin |
| A / 7 | pBS-F1-GAA | Ampicillin |
| A / 8 | pCS2_F1-GAC | Ampicillin |
| A / 9 | pCS2-F1-GAG-F2-GAG-F3-AAG | Ampicillin |
| A / 10 | pCS2-F2-GAA | Ampicillin |
| A / 11 | pCS2-F1-GCA | Ampicillin |
| A / 12 | pCS2-F1-GCC | Ampicillin |
| B / 1 | pCS2-F1-GCG | Ampicillin |
| B / 2 | pCS2-F1-GCT | Ampicillin |
| B / 3 | pCS2-F1-GGA-F2-GTG-F3-TTG | Ampicillin |
| B / 4 | pCS2-F1-GGC | Ampicillin |
| B / 5 | pBS-F1-GGG | Ampicillin |
| B / 6 | pCS2-F1-GGT-F3-CAG | Ampicillin |
| B / 7 | pBS-F1-GTA | Ampicillin |
| B / 8 | pBS-F1-GTC | Ampicillin |
| B / 9 | pCS2-F1-GTG-F2-GGT-F3-GAA | Ampicillin |
| B / 10 | pCS2_F1-GTT | Ampicillin |
| B / 11 | pBS-F1-TAG | Ampicillin |
| B / 12 | pCS2-F1-TGG-F3-GGG | Ampicillin |
| C / 1 | pBS-F1-TGT | Ampicillin |
| C / 2 | pBS-F1-TTG | Ampicillin |
| C / 3 | pBS-F2-AAG | Ampicillin |
| C / 4 | pCS2-F2-ACG | Ampicillin |
| C / 5 | pBS-F2-AGG | Ampicillin |
| C / 6 | pBS-F2-ATG | Ampicillin |
| C / 7 | pBS-F2-CAG | Ampicillin |
| C / 8 | pBS-F2-CGG | Ampicillin |
| C / 9 | pBS-F2-GAT | Ampicillin |
| C / 10 | pCS2-F2-GCA | Ampicillin |
| C / 11 | pBS-F2-GCC | Ampicillin |
| C / 12 | pCS2-F2-GCG | Ampicillin |
| D / 1 | pCS2-F2-GCT | Ampicillin |
| D / 2 | pBS-F2-GGA | Ampicillin |
| D / 3 | pCS2-F2-GGC | Ampicillin |
| D / 4 | pBS-F3-GAG | Ampicillin |
| D / 5 | pBS-F2-GTA | Ampicillin |
| D / 6 | pBS-F2-GTC | Ampicillin |
| D / 7 | pCS2-F2-GTT-F3-GGC | Ampicillin |
| D / 8 | pBS-F2-TAG | Ampicillin |
| D / 9 | pCS2-F2-TGG | Ampicillin |
| D / 10 | pBS-F2-TGT | Ampicillin |
| D / 11 | pBS-F2-TTG | Ampicillin |
| D / 12 | pBS-F3-ACG | Ampicillin |
| E / 1 | pBS-F3-AGG | Ampicillin |
| E / 2 | pBS-F3-ATG | Ampicillin |
| E / 3 | pBS-F3-CGG | Ampicillin |
| E / 4 | pBS-F3-GAC | Ampicillin |
| E / 5 | pCS2-F3-GAT | Ampicillin |
| E / 6 | pBS-F3-GCC | Ampicillin |
| E / 7 | pCS2-F3-GCT | Ampicillin |
| E / 8 | pBS-F3-GGA | Ampicillin |
| E / 9 | pCS2-F3-GGT | Ampicillin |
| E / 10 | pBS-F3-GTA | Ampicillin |
| E / 11 | pBS-F3-GTC | Ampicillin |
| E / 12 | pCS2-F3-GTG | Ampicillin |
| F / 1 | pBS-F3-GTT | Ampicillin |
| F / 2 | pBS-F3-TAG | Ampicillin |
| F / 3 | pBS-F3-TGG | Ampicillin |
| F / 4 | pCS2-F3-TGT | Ampicillin |
| F / 5 | pBS-F3-GCG | Ampicillin |
| F / 6 | pCS2-F2-GGG | Ampicillin |
| F / 7 | pBS-F1-GAT | Ampicillin |
| F / 8 | pCS2-F3-GCA | Ampicillin |
| F / 9 | pCS2-F1-AGA | Ampicillin |
| F / 10 | pBS-F2-AGA | Ampicillin |
| F / 11 | pBS-F3-AGA | Ampicillin |
| H / 1 | pCS2-HA-Kdrl-3p-FokI-KK | Ampicillin |
| H / 2 | pCS2-Flag-Kdrl-5p-FokI-EL | Ampicillin |