TALEN Plasmids and Kits

TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. View Addgene's TALEN Guide.

These collections and accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays.

TALEN Kits

TALEN Kit	Description	Depositing PI
FusX TALEN Assembly System	Contains 336 pFusX clones allowing for Golden Gate assembly of 15-16 RVD TALEN inserts in one cloning step rather than two.	Stephen Ekker
Golden Gate TALEN 2.0	Golden Gate cloning method. Validated in multiple organisms.	Dan Voytas Adam Bogdanove
Joung Lab TAL Effector Engineering Reagents	Assembly via serial ligation. Validated in zebrafish somatic cells.	Keith Joung
Zhang Lab TALE Toolbox	PCR/Golden Gate cloning method. Optimized for human expression.	Feng Zhang
LIC TAL Effector Assembly Kit	Assembly by ligation-independent cloning (LIC). Validated in human cells.	Veit Hornung
Musunuru/Cowan Lab TALEN Kit	Kit comprised of 834 plasmids of pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly.	Kiran Musunuru and Chad Cowan
Ekker Lab TALEN Kit	All possible tetramers (264) of RVDs, compatible with Golden Gate TALEN and TAL Effector Kit 2.0.	Stephen Ekker
Platinum Gate TALEN Kit	Platinum TALENs assembled by Golden Gate. Validated in several species.	Takashi Yamamoto

Additional Plasmids for Use with Addgene TALEN Kits

For Use With:	ID	Plasmid	Depositing PI	Description
Golden Gate TALEN 2.0	38142	RCIscript-GoldyTALEN	Daniel Carlson	The plasmids pC-GoldyTALEN and RCIsricpt-GoldyTALEN are designed as destination vectors for the Voytas lab Golden Gate TALEN kit. The GoldyTALEN scaffold is truncated at both the N and C terminus and induces mutation at rates much higher than the parental vectors. pC-GoldyTALEN directs expression of TALENs from a truncated CAGs promoter. RCIscript-GoldyTALEN is designed for in vitro synthesis of TALEN mRNAs. Both 5’ and 3’ Xenopus β-globin UTRs are included in the vector to enhance expression of the message. Both vectors utilize homodimeric FokI domains.
	38143	pC-GoldyTALEN
Golden Gate TALEN 2.0	36033	pTAL5-BB	Tom Ellis	Plasmids pTAL5-BB and pTAL6-BB are designed as alternative destination vectors for the Golden Gate TALEN kit (Bogdanove/Voytas) in order to generate TALORs (TAL Orthongal Repressors) that can be used to custom repress gene expression in yeast. TALORs consist of the DNA-binding domain of a TALE, along with strong nuclear localisation tags and repress transcription initiation when targeted to DNA sequences within core promoter regions. pTAL5-BB contains the GAL1 promoter, placing TALORs built into this vector under galactose-inducible expression. pTAL6-BB contains the TEF1 promoter, giving constitutive expression of TALORs built into this vector.
	36034	pTAL6-BB
Golden Gate TALEN 2.0	37275	pCS2TAL3-DD	David Grunwald	pCS2TAL3-DD and pCS2TALE3-RR are next generation TALEN backbone vectors designed for use with the Voytas Golden Gate TALEN Kit and are used in place of pTAL1, 2, 3, or 4. For both plasmids sequence positions 1214–2210 of pTAL3 were cloned into a pCS2 expression vector resulting in shorter N- and C-terminal tal protein segments (136AA and 63AA, respectfully). This next generation architecture has been shown to increase mutation induction when using TALENs. The FokI domains (DD, RR) used are obligate heterodimers that require cloning of left and right TALEN monomer proteins into opposite vectors.
	37276	pCS2TAL3-RR
	48636	pCS2TAL3-RRR
	48637	pCS2TAL3-DDD
Golden Gate TALEN 2.0	37184	pCAG-T7-TALEN(Sangamo)-Destination	Pawel Pelczar	pCAG-T7-TALEN(Sangamo)-Destination constructs were designed for optimal mammalian expression of Voytas Golden Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN expression is driven by the strong CAG promoter or can be achieved by in vitro mRNA synthesis from the T7 promoter. Truncations were introduced to the N- and C-terminus of the pTAL3 TALEN backbone, which were initially published by Sangamo BioSciences (N153AA, C63AA) and showed robust cleavage activity in several later studies. pCAG-T7-TALEN(Sangamo)-Destination vectors are available with homodimeric or enhanced heterodimeric (ELD, KKR mutations) FokI domains.
	40131	pCAG-T7-TALEN(Sangamo)-FokI-KKR-Destination
	40132	pCAG-T7-TALEN(Sangamo)-FokI-ELD-Destination
Golden Gate TALEN 2.0	1000000030	Yamamoto Lab TALEN Construction and Evaluation Accessory Pack	Takashi Yamamoto	This accessory pack contains modified pFUS array vectors, destination vectors and a reporter vector for mammalian cell-based validation assay that are designed for use with the Golden Gate TALEN and TAL Effector Kit. These modified pFUS vectors can reduce the number of module plasmids and improve the success rate of Golden Gate assembly. Destination vectors, pcDNA-TAL-NC2 and pCAGGS-TAL-NC2, are mammalian expression/mRNA synthesis vectors containing unique shorter N- and C-terminal domains (N153AA, C47AA). The reporter vector pGL4-SSA can be used for single-strand annealing assay, which enables the evaluation of TALEN activities in cultured cells.
Golden Gate TALEN 2.0	47388	pcDNA3.1-GoldenGate	Charles Gersbach	These new destination vectors can be used to create TALE-transcription factors and are used at the final assembly step in the Golden Gate protocol. Both plasmids 47388 and 47389 include a Flag epitope tag and an SV40 NLS at the N terminus, a 152-residue deletion from the N terminus of the wild-type TALE protein (previously shown to preserve the DNA-binding ability of TALEs), 63 wild-type TAL amino acids after the repeat domain, and C-terminal SV40 NLS and HA tags. Plasmid 47389 also contains the VP64 domain (four repeats of the minimal activation domain of VP16) at the C terminus.
	47389	pcDNA3.1-GoldenGate-VP64
Golden Gate TALEN 2.0	47874	pTALYM3	Maria-Elena Torres-Padilla	These plasmids were designed for mammalian expression of fluorescent TAL effectors to visualize subcellular positioning of target DNA sequences in living cells (TALE-mediated Genome Visualization [TGV]). pTALYM3 and pTALYM4 contain TALE fused with mClover and mRuby2, respectively, and can be used as destination vectors for Voytas Golden Gate-assembly kit.
	47875	pTALYM4
Golden Gate TALEN 2.0	48705	pTAL7a	Boris Greber	Application of TALEN technology in hPSCs pTAL7a and pTAL7b vectors were designed for use with the Voytas Lab Golden Gate TALEN kit, replacing pTAL4. The pTAL7 destination plasmids were optimized for TALEN expression in mammalian systems including hard-to-transfect cell lines. Features include (i) Esp3I (BsmBI) restriction sites for full compatibility with the Golden Gate TALEN kit, (ii) a lacZ fragment for blue/white-screening in E.coli, (iii) CAG promoter and Kozak sequence to drive efficient TALEN expression in mammalian cells, (iv) an improved, truncated TALE backbone architecture as established by Miller et al. (PMID: 21179091), as well as (v) distinct selection cassettes on pTAL7a and pTAL7b for enrichment of double-transfected mammalian cells. In addition, a T7 promoter was included for generating TALEN mRNAs through in vitro transcription as an alternative. Based on transient double-selection of transfected cells, the system has successfully been employed for deletion mutagenesis and gene targeting in human and mouse pluripotent stem cells.
	48706	pTAL7b
Joung Lab REAL Assembly TALEN Kit	49042	EMM65	Bradley Bernstein and Eric Mendenhall	The TALE plasmid vectors described on this page are designed to target and edit the epigenome of putative enhancers in mammalian cells. These plasmids allow for cloning DNA fragments encoding TAL effector repeat arrays (assembled using the REAL, REAL-Fast, or FLASH methods) for expression as TALE-LSD1 fusions. It was recently shown that TALE fusions with the histone demethylase LSD1 (Lysine specific demethylase) made using these plasmids along with the FLASH method can decrease the amount of Histone H3 lysine 4 (H3K4) di-methylation at the target locus (Mendenhall et al., 2013). This epigenome editing was also shown to alter regulation of nearby gene expression. The vectors listed below have the EF1α promoter for efficient mammalian cell expression, and encode one of three 0.5 TALE repeats (C, T, and G) and the full length human LSD1 at the carboxy-terminus of the protein.
	49043	EMM67
	49044	EMM71T
Golden Gate TALEN 2.0	49401	pBlue-TAL	Michal Zurovec	pBlue-TAL was designed for use with the Voytas Lab Golden Gate TALEN kit. This backbone is suitable for the TALEN-based generation of germline mutations in Bombyx mori and Drosophila melanogaster.
Zhang Lab TALE Toolbox	49622 - 49647	24 plasmids	Thoru Pederson	TALEColor plasmids: destination vectors for use in labeling specific repetitive DNA sequences in human chromosomes