Mutagenesis mediated by CRISPR/Cas9 in the red imported fire ant, Solenopsis invicta
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Insectes Sociaux
TECHNICAL ARTICLE
Mutagenesis mediated by CRISPR/Cas9 in the red imported fire ant, Solenopsis invicta Y.‑K. Chiu1 · J.‑C. Hsu1 · T. Chang1 · Y.‑C. Huang1 · J. Wang1 Received: 4 October 2019 / Revised: 14 December 2019 / Accepted: 7 February 2020 © International Union for the Study of Social Insects (IUSSI) 2020
Abstract CRISPR/Cas9 mediated mutagenesis has revolutionized the testing of gene function in both model and non-model organisms. The red imported fire ant, Solenopsis invicta, is the best-studied ant species because of their painful sting, aggressive nature, and their detrimental effects on invaded ecosystems. We have developed a microinjection protocol for CRISPR/Cas9 mutagenesis of fire ant embryos. As a proof-of-principle we generated individuals mosaic for the knockout of Sinv-spitz or Gp-9. We verified that many injected individuals carry mutations, often to high frequency of the cells within the individual. Observations at the first instar larval stage indicate that mutating Sinv-spitz does not produce a loss of oenocyte phenotype, as would have been expected based on the homologous function of the fruit fly, Drosophila melanogaster, spitz. Our success indicates that CRISPR/Cas9 mutagenesis should be a useful technique for studying gene function in the fire ant at the individual and possibly social levels. Keywords Microinjection · Mutagenesis · CRISPR/Cas9 · Oenocyte · Fire ant
Introduction The discovery of the CRISPR/Cas9 genome editing system and its application for genome manipulation has greatly expanded the set of organisms for which functional tests are now possible (Adli 2018; Cong et al. 2013; Hsu et al. 2014; Jinek et al. 2012; Wiedenheft et al. 2012). Ants and other social insects have been beloved models for social and evolutionary biologists. They also have been poorly studied at the molecular-genetic level partly because of the Yu-Kun Chiu, Jung-Chen Hsu, and Tiffany Chang contributed equally. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00040-020-00755-8) contains supplementary material, which is available to authorized users. * Y.‑C. Huang [email protected] * J. Wang [email protected] 1
Biodiversity Research Center, Academia Sinica, Taipei, Taiwan
Present Address: CH Biotech R&D Co., Ltd., Nantou City, Taiwan
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lack of satisfactory genetic tools, with the exception of the honeybee (Cridge et al. 2017; Kohno et al. 2016; Li-Byarlay 2013; Rittschof and Robinson 2016; Roth et al. 2019; Schulte et al. 2014). This may be changing for the ants, as recently CRISPR/Cas9 mediated mutagenesis was successfully applied to two ant species, Ooceraea biroi and Harpegnathos saltator (Trible et al. 2017; Yan et al. 2017). The development of CRISPR/Cas9 genome editing techniques for other ants would also be extremely useful. The red imported fire ant, Solenopsis invicta, is a wellstudied ant. It is a major pest species of important economic and ecological impact (Tschinkel 2006). Additionally,
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