title: Engineering Customized TALENs Using the Platinum Gate TALEN Kit.
abstract: Among various strategies for constructing customized transcription activator-like effector nucleases (TALENs), the Golden Gate assembly is the most widely used and most characterized method. The principle of Golden Gate assembly involves cycling reactions of digestion and ligation of multiple plasmids in a single tube, resulting in PCR-, fragmentation-, and purification-free concatemerization of DNA-binding repeats. Here, we describe the protocols for Golden Gate assembly-based TALEN construction using the Platinum Gate TALEN Kit, which allows generation of highly active Platinum TALENs.
authors.affiliation: Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8526, Japan.
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Sakuma
authors.affiliation: Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8526, Japan. tybig@hiroshima-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamoto
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D004720:Endonucleases / Q000096:biosynthesis / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D015534:Trans-Activators; D004720:Endonucleases
keywords: 4-Module assembly; Golden Gate assembly; Modular assembly; Platinum Gate TALEN Kit; Platinum TALEN