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Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes

Author

Listed:
  • Xuechun Feng

    (University of California San Diego)

  • Víctor López Del Amo

    (University of California San Diego)

  • Enzo Mameli

    (Harvard Medical School
    Boston University, School of Medicine)

  • Megan Lee

    (University of California San Diego)

  • Alena L. Bishop

    (University of California San Diego)

  • Norbert Perrimon

    (Harvard Medical School
    HHMI, Harvard Medical School)

  • Valentino M. Gantz

    (University of California San Diego)

Abstract

Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, and avian malaria, posing a constant threat to public health, livestock, companion animals, and endangered birds. While rising insecticide resistance has threatened the control of Culex mosquitoes, advances in CRISPR genome-editing tools have fostered the development of alternative genetic strategies such as gene drive systems to fight disease vectors. However, though gene-drive technology has quickly progressed in other mosquitoes, advances have been lacking in Culex. Here, we develop a Culex-specific Cas9/gRNA expression toolkit and use site-directed homology-based transgenesis to generate and validate a Culex quinquefasciatus Cas9-expressing line. We show that gRNA scaffold variants improve transgenesis efficiency in both Culex quinquefasciatus and Drosophila melanogaster and boost gene-drive performance in the fruit fly. These findings support future technology development to control Culex mosquitoes and provide valuable insight for improving these tools in other species.

Suggested Citation

  • Xuechun Feng & Víctor López Del Amo & Enzo Mameli & Megan Lee & Alena L. Bishop & Norbert Perrimon & Valentino M. Gantz, 2021. "Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes," Nature Communications, Nature, vol. 12(1), pages 1-13, December.
  • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-23239-0
    DOI: 10.1038/s41467-021-23239-0
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    Cited by:

    1. Raghuvir Viswanatha & Enzo Mameli & Jonathan Rodiger & Pierre Merckaert & Fabiana Feitosa-Suntheimer & Tonya M. Colpitts & Stephanie E. Mohr & Yanhui Hu & Norbert Perrimon, 2021. "Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos," Nature Communications, Nature, vol. 12(1), pages 1-13, December.
    2. Tim Harvey-Samuel & Xuechun Feng & Emily M. Okamoto & Deepak-Kumar Purusothaman & Philip T. Leftwich & Luke Alphey & Valentino M. Gantz, 2023. "CRISPR-based gene drives generate super-Mendelian inheritance in the disease vector Culex quinquefasciatus," Nature Communications, Nature, vol. 14(1), pages 1-12, December.
    3. Sara Sanz Juste & Emily M. Okamoto & Christina Nguyen & Xuechun Feng & Víctor López Del Amo, 2023. "Next-generation CRISPR gene-drive systems using Cas12a nuclease," Nature Communications, Nature, vol. 14(1), pages 1-8, December.

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