title: Transcription activator-like effector nuclease (TALEN)-mediated female-specific sterility in the silkworm, Bombyx mori.
abstract: Engineering sex-specific sterility is critical for developing transgene-based sterile insect technology. Targeted genome engineering achieved by customized zinc-finger nuclease, transcription activator-like effector nuclease (TALEN) or clustered, regularly interspaced, short palindromic repeats/Cas9 systems has been exploited extensively in a variety of model organisms; however, screening mutated individuals without a detectable phenotype is still challenging. In addition, genetically recessive mutations only detectable in homozygotes make the experiments time-consuming. In the present study, we model a novel genetic system in the silkworm, Bombyx mori, that results in female-specific sterility by combining transgenesis with TALEN technologies. This system induces sex-specific sterility at a high efficiency by targeting the female-specific exon of the B. mori doublesex (Bmdsx) gene, which has sex-specific splicing isoforms regulating somatic sexual development. Transgenic animals co-expressing TALEN left and right arms targeting the female-specific Bmdsx exon resulted in somatic mutations and female mutants lost fecundity because of lack of egg storage and abnormal external genitalia. The wild-type sexual dimorphism of abdominal segment was not evident in mutant females. In contrast, there were no deleterious effects in mutant male moths. The current somatic TALEN technologies provide a promising approach for future insect functional genetics, thus providing the basis for the development of attractive genetic alternatives for insect population management.
authors.affiliation: Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China.
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.forename: L
authors.initials: L
authors.lastname: Ling
authors.forename: B
authors.initials: B
authors.lastname: Zeng
authors.forename: A A
authors.initials: AA
authors.lastname: James
authors.forename: A
authors.initials: A
authors.lastname: Tan
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D012831:Bombyx / Q000196:embryology / Q000235:genetics*; D005260:Female; D005818:Genetic Engineering / Q000379:methods*; D005836:Genitalia, Female / Q000002:abnormalities; D049750:Genome, Insect / Q000235:genetics; D007247:Infertility, Female / Q000235:genetics*; D007300:Insect Control / Q000379:methods; D008297:Male; D016296:Mutagenesis; D020033:Protein Isoforms; D012726:Sexual Behavior, Animal; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation; D019076:Transgenes
chemical_list: D020033:Protein Isoforms; D014157:Transcription Factors
keywords: doublesex; gene targeting; genitalia; morphology