Expanding the CRISPR toolbox in Culicine mosquitoes: in vitro validation of Pol III promoters

Michelle Anderson, Jessica Purcell, Sebald Verkuijl, Victoria Norman, Philip Leftwich, Tim Harvey-Samuel, Luke Alphey

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19 Citations (Scopus)
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CRISPR–Cas9-based “gene drive” technologies have been proposed as a novel and effective means of controlling human diseases vectored by mosquitoes. However, more complex designs than those demonstrated to date—and an expanded molecular toolbox with which to build them—will be required to overcome the issues of resistance formation/evolution and drive spatial/temporal limitation. Foreseeing this need, we assessed the sgRNA transcriptional activities of 33 phylogenetically diverse insect Polymerase III promoters using three disease-relevant Culicine mosquito cell lines (Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus). We show that U6 promoters work across species with a range of transcriptional activity levels and find 7SK promoters to be especially promising because of their broad phylogenetic activity. We further show that U6 promoters can be substantially truncated without affecting transcriptional levels. These results will be of great utility to researchers involved in developing the next generation of gene drives.
Original languageEnglish
Pages (from-to)678-681
Number of pages4
JournalACS Synthetic Biology
Issue number3
Early online date4 Mar 2020
Publication statusPublished - 20 Mar 2020


  • 7SK promoter
  • Cas9
  • Polymerase III
  • U6 promoter
  • gene drive
  • mosquito

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