IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v12y2021i1d10.1038_s41467-021-27129-3.html
   My bibliography  Save this article

Bioinformatic and cell-based tools for pooled CRISPR knockout screening in mosquitos

Author

Listed:
  • Raghuvir Viswanatha

    (Blavatnik Institute, Harvard Medical School)

  • Enzo Mameli

    (Blavatnik Institute, Harvard Medical School
    Boston University School of Medicine, 620 Albany Street)

  • Jonathan Rodiger

    (Blavatnik Institute, Harvard Medical School)

  • Pierre Merckaert

    (Blavatnik Institute, Harvard Medical School)

  • Fabiana Feitosa-Suntheimer

    (Boston University School of Medicine, 620 Albany Street)

  • Tonya M. Colpitts

    (Boston University School of Medicine, 620 Albany Street)

  • Stephanie E. Mohr

    (Blavatnik Institute, Harvard Medical School)

  • Yanhui Hu

    (Blavatnik Institute, Harvard Medical School)

  • Norbert Perrimon

    (Blavatnik Institute, Harvard Medical School
    HHMI, Harvard Medical School)

Abstract

Mosquito-borne diseases present a worldwide public health burden. Current efforts to understand and counteract them have been aided by the use of cultured mosquito cells. Moreover, application in mammalian cells of forward genetic approaches such as CRISPR screens have identified essential genes and genes required for host-pathogen interactions, and in general, aided in functional annotation of genes. An equivalent approach for genetic screening of mosquito cell lines has been lacking. To develop such an approach, we design a new bioinformatic portal for sgRNA library design in several mosquito genomes, engineer mosquito cell lines to express Cas9 and accept sgRNA at scale, and identify optimal promoters for sgRNA expression in several mosquito species. We then optimize a recombination-mediated cassette exchange system to deliver CRISPR sgRNA and perform pooled CRISPR screens in an Anopheles cell line. Altogether, we provide a platform for high-throughput genome-scale screening in cell lines from disease vector species.

Suggested Citation

  • 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.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-27129-3
    DOI: 10.1038/s41467-021-27129-3
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-021-27129-3
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-021-27129-3?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. October M. Sessions & Nicholas J. Barrows & Jayme A. Souza-Neto & Timothy J. Robinson & Christine L. Hershey & Mary A. Rodgers & Jose L. Ramirez & George Dimopoulos & Priscilla L. Yang & James L. Pear, 2009. "Discovery of insect and human dengue virus host factors," Nature, Nature, vol. 458(7241), pages 1047-1050, April.
    2. 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.
    Full references (including those not matched with items on IDEAS)

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. 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.
    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. Linhui Hao & Qiuling He & Zhishi Wang & Mark Craven & Michael A Newton & Paul Ahlquist, 2013. "Limited Agreement of Independent RNAi Screens for Virus-Required Host Genes Owes More to False-Negative than False-Positive Factors," PLOS Computational Biology, Public Library of Science, vol. 9(9), pages 1-20, September.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-27129-3. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.