Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases

Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Hadley W. Horch, Naoya Hamaguchi, Taro Nakamura, Tetsuya Bando, Hideyo Ohuchi, Takashi Yamamoto, Sumihare Noji, Taro Mito

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically relatively basal and comprise many pests. However, the absence of a sophisticated genetic model system, or targeted gene-manipulation system, has limited research on hemimetabolous species. Here we use zinc-finger nuclease and transcription activator-like effector nuclease technologies to produce genetic knockouts in the hemimetabolous insect Gryllus bimaculatus. Following the microinjection of mRNAs encoding zinc-finger nucleases or transcription activator-like effector nucleases into cricket embryos, targeting of a transgene or endogenous gene results in sequence-specific mutations. Up to 48% of founder animals transmit disrupted gene alleles after zinc-finger nucleases microinjection compared with 17% after microinjection of transcription activator-like effector nucleases. Heterozygous offspring is selected using mutation detection assays that use a Surveyor (Cel-I) nuclease, and subsequent sibling crosses create homozygous knockout crickets. This approach is independent from a mutant phenotype or the genetic tractability of the organism of interest and can potentially be applied to manage insect pests using a non-transgenic strategy.

Original languageEnglish
Article number1017
JournalNature Communications
Volume3
DOIs
Publication statusPublished - 2012
Externally publishedYes

Fingerprint

nuclease
effectors
insects
genome
Zinc Fingers
Microinjections
Transcription
Insects
Gryllidae
Zinc
Genes
zinc
Genome
crickets
Mutation
genes
Genetic Models
Transgenes
mutations
Assays

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

Watanabe, T., Ochiai, H., Sakuma, T., Horch, H. W., Hamaguchi, N., Nakamura, T., ... Mito, T. (2012). Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases. Nature Communications, 3, [1017]. https://doi.org/10.1038/ncomms2020

Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases. / Watanabe, Takahito; Ochiai, Hiroshi; Sakuma, Tetsushi; Horch, Hadley W.; Hamaguchi, Naoya; Nakamura, Taro; Bando, Tetsuya; Ohuchi, Hideyo; Yamamoto, Takashi; Noji, Sumihare; Mito, Taro.

In: Nature Communications, Vol. 3, 1017, 2012.

Research output: Contribution to journalArticle

Watanabe, T, Ochiai, H, Sakuma, T, Horch, HW, Hamaguchi, N, Nakamura, T, Bando, T, Ohuchi, H, Yamamoto, T, Noji, S & Mito, T 2012, 'Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases', Nature Communications, vol. 3, 1017. https://doi.org/10.1038/ncomms2020
Watanabe, Takahito ; Ochiai, Hiroshi ; Sakuma, Tetsushi ; Horch, Hadley W. ; Hamaguchi, Naoya ; Nakamura, Taro ; Bando, Tetsuya ; Ohuchi, Hideyo ; Yamamoto, Takashi ; Noji, Sumihare ; Mito, Taro. / Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases. In: Nature Communications. 2012 ; Vol. 3.
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