REAL Assembly TALEN Kit
(Kit #
1000000017
)
Depositing Lab: Keith Joung
This 32 plasmid TALEN kit is used to assemble any combination of TALE repeats into arrays of any desired length using the REAL, REAL-Fast or FLASH assembly methods described by the Joung lab.
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
Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Sanders JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung KJ, Yeh JRJ. Nat Biotechnol. 2011 Aug 5;29(8):697-8.PubMed PMID 21822241
Description
The Joung Lab has described a rapid and robust protocol for constructing TALE repeat arrays of any desired sequence, referred to as the REAL (Restriction Enzyme And Ligation) assembly method (Sander et al., Nat Biotechnol. 2011) as well as REAL-Fast and FLASH methods. The Joung lab TALEN kit utilizes a set of 28 plasmids each encoding a single TALE repeat domain and flanked by restriction sites used for ligation into longer arrays and 4 different TALEN expression vectors. DNA fragments encoding TALE repeat arrays can be easily transferred by simple cloning into one of four different TALEN expression vectors (each possessing one of the four different carboxy-terminal 0.5 TALE repeat domains). Using REAL, REAL-Fast or FLASH assemnbly methods, any combination of TALE repeats can be assembled into arrays of any desired length.
Kit Documentation
Joung lab TALEN kit.pdf 368.2Â KBResources
More detailed information about TALE assembly, including detailed and up-to-date protocols, can be found on the TALengineering website. Users may also wish to join the TALengineering newsgroup
An upgraded version of the ZiFiT Targeter program is available to enable users to identify both dimeric TALEN and monomeric TALE transcriptional activator target sites and to simplify the construction of TALEN expression plasmids using REAL, REAL-Fast or FLASH assembly. This new version of ZiFiT Targeter enables users to search their sequences of interest for potential TALEN or TALE transcriptional activator target sites and then generates individual user-friendly graphical roadmaps for each target that guide the user through the REAL or REAL-Fast assembly process or lists of plasmid ID numbers required for FLASH assembly. The software is publicly available without registration at: http://zifit.partners.org.
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 TALEN kit used for TALE assembly was a gift from Keith Joung (Addgene kit # 1000000017)"
For your Reference section:
Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Sanders JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung KJ, Yeh JRJ. Nat Biotechnol. 2011 Aug 5;29(8):697-8. PubMed PMID 21822241
Joung Lab REAL Assembly TALEN Kit - #1000000017
- 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 | TAL006 | Ampicillin |
| A / 2 | TAL007 | Ampicillin |
| A / 3 | TAL009 | Ampicillin |
| A / 4 | TAL010 | Ampicillin |
| A / 5 | TAL011 | Ampicillin |
| A / 6 | TAL012 | Ampicillin |
| A / 7 | TAL014 | Ampicillin |
| A / 8 | TAL015 | Ampicillin |
| A / 9 | TAL016 | Ampicillin |
| A / 10 | TAL017 | Ampicillin |
| A / 11 | TAL019 | Ampicillin |
| A / 12 | TAL020 | Ampicillin |
| B / 1 | TAL021 | Ampicillin |
| B / 2 | TAL022 | Ampicillin |
| B / 3 | TAL024 | Ampicillin |
| B / 4 | TAL025 | Ampicillin |
| B / 5 | TAL026 | Ampicillin |
| B / 6 | TAL027 | Ampicillin |
| B / 7 | TAL029 | Ampicillin |
| B / 8 | TAL030 | Ampicillin |
| B / 9 | TAL031 | Ampicillin |
| B / 10 | TAL032 | Ampicillin |
| B / 11 | TAL034 | Ampicillin |
| B / 12 | TAL035 | Ampicillin |
| C / 1 | TAL036 | Ampicillin |
| C / 2 | TAL037 | Ampicillin |
| C / 3 | TAL039 | Ampicillin |
| C / 4 | TAL040 | Ampicillin |
| C / 5 | JDS70 | Ampicillin |
| C / 6 | JDS71 | Ampicillin |
| C / 7 | JDS74 | Ampicillin |
| C / 8 | JDS78 | Ampicillin |