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A new technology for genome editing may put the zinc finger nuclease franchise out of business, some believe. Not so fast, say the finger people. Laura DeFrancesco reports. 
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"Zinc finger nucleases have been used to generate DSBs and subsequently, for genome editing but with low efficiency and reproducibility. ... Here, we report the generation of a Hax3-based hybrid TALE nuclease with a user-selected DNA binding specificity. ... Transient expression assays in tobacco leaves suggest that the hybrid nuclease creates DSB (double strand breaks) in its target sequence, which is subsequently repaired by nonhomologous end-joining repair. ... great potential for site-specific genome modification in plants and eukaryotes in general"
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It is this repeat domain that is responsible for specific DNA sequence recognition. Each repeat is almost identical with the exception of two variable amino acids termed the repeat-variable diresidues.
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"one needs only to modify the" two variable amino acids per repeat, called "RVDs, to generate designer TALEs"
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The most complete review on TAL effectors from Xanthomonas bacteria to date
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"Additionally, TALE-NT provides tools to design single TAL effectors to bind to a DNA sequence, and to identify binding sites in a DNA sequence for a given TAL effector.
The TALE-NT website is maintained by the Bogdanove laboratory at Iowa State University."
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From
jb
26 TAL effector genes in newly sequenced Xanthomonas oryzae pv. oryzicola strain BLS256, none in Xanthomonas campestris raphani 756C
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A "Golden TAL Technology" to assemble custom DNA binding domains and their use es activators in human cells
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The other paper describing the TAL effector code including its powerful application to design artifical DNA binding domains
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From
issuu
This article describes how the DNA binding specificity of TAL effectors was discovered
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"Os11N3 is directly upregulated by TalC and identify a TalC DNA target box within the Os11N3 upstream sequence"
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A TAL effector of "Xanthomonas oryzae pv oryzae (Xoo) transcriptionally activates rice (Oryza sativa) susceptibility gene Xa13 to cause bacterial blight disease. Xa13 encodes an indispensable plasma membrane protein"..."We show that the XA13 protein cooperates with two other proteins"..."to promote removal of copper from xylem vessels, where Xoo multiplies and spreads to cause disease"
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The TAL Effector Technology license was jointly signed with inventors Jens Boch, Ulla Bonas, Thomas Lahaye and Sebastian Schornack, and the Two Blades Foundation (2Blades), which exclusively licensed plant research tool ...
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Another publication describing the use of type IIS restriction enzymes in two sequential cut-ligase reactions to build TAL DNA binding domain repeat arrays
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"Recent elucidation of the basis for specificity in DNA binding by TALEs expedites further discovery and opens the door to biotechnological applications. This article reviews the most significant findings in TALE research, with a focus on recent advances, and discusses future prospects including pressing questions yet to be answered"
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TAL effector - Description: TAL effectors (TALEs) are proteins secreted by Xanthomonas bacteria via their type III secretion system when they infect various plant species.
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Transcription activator-like effector nucleases (TALENs) are a new technology for modifying the genome at specific loci of interest. Hockemeyer et al. now demonstrate the utility of TALENs for gene targeting in human pluripotent stem cells.
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"We leverage codon degeneracy and type IIs restriction enzymes to generate orthogonal ligation linkers between individual repeat monomers, thus allowing full-length, customized, repeat domains to be constructed by hierarchical ligation. We" ... "demonstrate that they can specifically modulate transcription of endogenous genes (SOX2 and KLF4) in human cells"
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One of two papers describing the original discovery of a linear code for DNA-binding site prediction
Image: Adam Bogdanove and Harry Horner/Iowa State University; false color: Yael Kats/Science
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A TAL effector is a protein of bacterial origin and is able to bind to DNA sequences. The bound sequence is defined by the structure of the TAL effector protein. Manipulate the effector protein and you can direct to any binding site you want. Powerful biotechnology, I say...
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