title: Ligation-independent cloning (LIC) assembly of TALEN genes.
abstract: Modular DNA binding protein architectures hold the promise of wide application in functional genomic studies. Functionalization of DNA binding proteins, e.g. using the FokI nuclease domain, provides a potent tool to induce DNA double strand breaks at user-defined genomic loci. In this regard, TAL (transcription activator-like) effector proteins, secreted by bacteria of the Xanthomonas family, provide the highest degree of modularity in their DNA binding mode. However, the assembly of large and highly repetitive TALE protein coding genes can be challenging. We describe a ligation-independent cloning (LIC) based method to allow high-throughput assembly of TALE nuclease genes at high fidelity and low effort and cost.
authors.affiliation: Institute of Molecular Medicine, University Hospital, University of Bonn, 53127, Bonn, Germany.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Schmid-Burgk
authors.forename: Tobias
authors.initials: T
authors.lastname: Schmidt
authors.forename: Veit
authors.initials: V
authors.lastname: Hornung
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism; D018390:Gene Targeting; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D059765:Homologous Recombination; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D015202:Protein Engineering / Q000379:methods
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D004720:Endonucleases