title: Gene knockout by targeted mutagenesis in a hemimetabolous insect, the two-spotted cricket Gryllus bimaculatus, using TALENs.
abstract: Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically basal. These insects include many deleterious species. The cricket, Gryllus bimaculatus, is an emerging model for hemimetabolous insects, based on the success of RNA interference (RNAi)-based gene-functional analyses and transgenic technology. Taking advantage of genome-editing technologies in this species would greatly promote functional genomics studies. Genome editing using transcription activator-like effector nucleases (TALENs) has proven to be an effective method for site-specific genome manipulation in various species. TALENs are artificial nucleases that are capable of inducing DNA double-strand breaks into specified target sequences. Here, we describe a protocol for TALEN-based gene knockout in G. bimaculatus, including a mutant selection scheme via mutation detection assays, for generating homozygous knockout organisms.
authors.affiliation: Department of Life Systems, Institute of Technology and Science, The University of Tokushima, 2-1 Minami-Jyosanjima-cho, Tokushima 770-0815, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Department of Life Systems, Institute of Technology and Science, The University of Tokushima, 2-1 Minami-Jyosanjima-cho, Tokushima 770-0815, Japan.
authors.forename: Sumihare
authors.initials: S
authors.lastname: Noji
authors.affiliation: Department of Life Systems, Institute of Technology and Science, The University of Tokushima, 2-1 Minami-Jyosanjima-cho, Tokushima 770-0815, Japan. Electronic address: mito@bio.tokushima-u.ac.jp.
authors.forename: Taro
authors.initials: T
authors.lastname: Mito
mesh_terms: D000818:Animals; D003851:Deoxyribonucleases / Q000737:chemistry; D055786:Gene Knockout Techniques; D006135:Gryllidae / Q000235:genetics*; D006720:Homozygote; D016297:Mutagenesis, Site-Directed
chemical_list: D003851:Deoxyribonucleases
keywords: Cricket; Homozygous knockout; TALEN; Targeted mutagenesis