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LIC TAL Effector Assembly Kit
(Kit # 1000000023 )

Depositing Lab:   Veit Hornung

This kit provides a LIC (ligation-independent cloning)-ready TALE repeat unit library of 2-mer fragments that can be assembled into an expression ready TALEN construct in 2 hierarchical assembly steps.

This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80oC immediately upon arrival.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

A ligation-independent cloning technique for high-throughput assembly of transcription activator-like effector genes. Schmid-Burgk JL, Schmidt T, Kaiser V, Höning K, Hornung V. Nat Biotechnol 2012 Dec 16. doi: 10.1038/nbt.2460. PubMed PMID 23242165.

Description

Transcription activator-like effector (TALE) proteins derived from Xanthomonas species have emerged as powerful scaffolds for engineering DNA binding proteins of user defined specificity. Their DNA recognition is governed by an array of modular repeat units in which two hypervariable amino acids, known as the repeat variable di-residue (RVD), determine binding to a specific nucleobase. The repeat units detect their target bases in a largely context-independent manner, which allows the generation of TALE proteins of de novo specificity based on the recently deciphered RVD - nucleobase code. Synthetic TALE proteins can be functionalized with a nuclease domain, allowing facile editing of eukaryotic genomic loci.

The Hornung lab has developed a simple high-fidelity ligation-independent cloning (LIC) technique that enables the semi‐automated generation of TALE genes at high throughput merely requiring the combinatorial mixing of fluids. To this effect, they generated a LIC-ready TALE repeat unit library of 2-mer fragments that can be assembled into an expression ready TALEN construct in 2 hierarchical assembly steps. In the first assembly reaction three 2-mer fragments are assembled into a 6-mer fragment, of which three fragments are assembled into an 18-mer in the second reaction. The 18-mer fragment will already be contained in an expression ready backbone with the last half repeat unit and a C-terminal nuclease domain, giving rise to an 18.5 RVD TALEN construct.

Here is a short movie describing the LIC assembly technology:

The whole set of 2-mer fragments and the corresponding backbones are available in a 96-well plate. We recommend generating a pre-digested and pre-chewed fragment library for all subsequent assembly reactions based on the layout of the library plate. The pipetting directions obtained from the assembly calculator tool are based on these coordinates.

Kit Documentation

Icon LIC TAL Kit (1.8 MB)

Protocols

Icon LIC TALEN Protocols (132.1 KB)

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 TALENs was a gift from Veit Hornung (Addgene kit # 1000000023)"

For your Reference section:

A ligation-independent cloning technique for high-throughput assembly of transcription activator-like effector genes. Schmid-Burgk JL, Schmidt T, Kaiser V, Höning K, Hornung V. Nat Biotechnol 2012 Dec 16. doi: 10.1038/nbt.2460. PubMed PMID 23242165.

LIC TAL Effector Assembly Kit - #1000000023

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. 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

Inventory

Well Plasmid Resistance
A / 1 TAL-1AA2
Ampicillin
A / 2 TAL-1AC2
Ampicillin
A / 3 TAL-1AG2
Ampicillin
A / 4 TAL-1AT2
Ampicillin
A / 5 TAL-1CA2
Ampicillin
A / 6 TAL-1CC2
Ampicillin
A / 7 TAL-1CG2
Ampicillin
A / 8 TAL-1CT2
Ampicillin
A / 9 TAL-1GA2
Ampicillin
A / 10 TAL-1GC2
Ampicillin
A / 11 TAL-1GG2
Ampicillin
A / 12 TAL-1GT2
Ampicillin
B / 1 TAL-1TA2
Ampicillin
B / 2 TAL-1TC2
Ampicillin
B / 3 TAL-1TG2
Ampicillin
B / 4 TAL-1TT2
Ampicillin
B / 5 TAL-3AA4
Ampicillin
B / 6 TAL-3AC4
Ampicillin
B / 7 TAL-3AG4
Ampicillin
B / 8 TAL-3AT4
Ampicillin
B / 9 TAL-3CA4
Ampicillin
B / 10 TAL-3CC4
Ampicillin
B / 11 TAL-3CG4
Ampicillin
B / 12 TAL-3CT4
Ampicillin
C / 1 TAL-3GA4
Ampicillin
C / 2 TAL-3GC4
Ampicillin
C / 3 TAL-3GG4
Ampicillin
C / 4 TAL-3GT4
Ampicillin
C / 5 TAL-3TA4
Ampicillin
C / 6 TAL-3TC4
Ampicillin
C / 7 TAL-3TG4
Ampicillin
C / 8 TAL-3TT4
Ampicillin
C / 9 TALEN-ID12-A
Ampicillin
C / 10 TALEN-ID12-C
Ampicillin
C / 11 TALEN-ID12-G
Ampicillin
C / 12 TALEN-ID12-T
Ampicillin
D / 1 TAL-2AA3
Ampicillin
D / 2 TAL-2AC3
Ampicillin
D / 3 TAL-2AG3
Ampicillin
D / 4 TAL-2AT3
Ampicillin
D / 5 TAL-2CA3
Ampicillin
D / 6 TAL-2CC3
Ampicillin
D / 7 TAL-2CG3
Ampicillin
D / 8 TAL-2CT3
Ampicillin
D / 9 TAL-2GA3
Ampicillin
D / 10 TAL-2GC3
Ampicillin
D / 11 TAL-2GG3
Ampicillin
D / 12 TAL-2GT3
Ampicillin
E / 1 TAL-2TA3
Ampicillin
E / 2 TAL-2TC3
Ampicillin
E / 3 TAL-2TG3
Ampicillin
E / 4 TAL-2TT3
Ampicillin
E / 5 TAL-4AA1
Ampicillin
E / 6 TAL-4AC1
Ampicillin
E / 7 TAL-4AG1
Ampicillin
E / 8 TAL-4AT1
Ampicillin
E / 9 TAL-4CA1
Ampicillin
E / 10 TAL-4CC1
Ampicillin
E / 11 TAL-4CG1
Ampicillin
E / 12 TAL-4CT1
Ampicillin
F / 1 TAL-4GA1
Ampicillin
F / 2 TAL-4GC1
Ampicillin
F / 3 TAL-4GG1
Ampicillin
F / 4 TAL-4GT1
Ampicillin
F / 5 TAL-4TA1
Ampicillin
F / 6 TAL-4TC1
Ampicillin
F / 7 TAL-4TG1
Ampicillin
F / 8 TAL-4TT1
Ampicillin
F / 9 TAL-4A1
Ampicillin
F / 10 TAL-4C1
Ampicillin
F / 11 TAL-4G1
Ampicillin
F / 12 TAL-4T1
Ampicillin
G / 1 TAL-BBL1-ID14
Kanamycin
G / 2 TAL-BBL1-ID43
Kanamycin
G / 3 TAL-BBL1-ID32
Kanamycin
G / 4 TAL-BBL1-ID21
Kanamycin
Data calculated @ 2017-11-24

Kit Plate Map - #1000000023

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