02928naa a2200313 a 450000100080000000500110000800800410001902200470006002400270010710000140013424501120014826000090026050006730026952012670094265300160220965300190222565300400224465300170228465300560230165300140235765300410237165300160241270000150242870000220244370000150246570000170248070000170249777301000251410643562023-10-24       2023    bl uuuu     u00u1 u    #d  a0962-1075 (print); 1365-2583 (electronic).7 a10.1111/imb.128752DOI1 aNOVAS, R.  aIdentification and functional analysis of Cochliomyia hominivorax U6 gene promoters.h[electronic resource]  c2023  aArticle history: Received 9 March 2023, Accepted 4 August 2023, First published 21 September 2023. --  Scott, M.J.; Department of Entomology and Plant Pathology, North Carolina State University, Campus Box 7613, Raleigh, NC, United States; email:mjscott3@ncsu.edu  -- FUNDING: This research was supported by an agreement between The Institut Pasteur de Montevideo and North Carolina State University and grants from the Inter-American Development Bank (IBD UR-T1227) and from INIA (FTPA N°359). Tatiana Basika, Rossina Novas, Pablo Fresia and Alejo Menchaca are members of SNI (National Research System, Uruguay). -- Document type: Article Hybrid Gold Open Access. --  aThe New World screwworm, Cochliomyia hominivorax, is an obligate parasite, which is a major pest of livestock. While the sterile insect technique was used very successfully to eradicate C. hominivorax from North and Central America, more cost-effective genetic methods will likely be needed in South America. The recent development of CRISPR/Cas9-based genetic approaches, such as homing gene drive, could provide a very efficient means for the suppression of C. hominivorax populations. One component of a drive system is the guide RNA(s) driven by a U6 gene promoter. Here, we have developed an in vivo assay to evaluate the activity of the promoters from seven C. hominivorax U6 genes. Embryos from the related blowfly Lucilia cuprina were injected with plasmid DNA containing a U6-promoter-guide RNA construct and a source of Cas9, either protein or plasmid DNA. Activity was assessed by the number of site-specific mutations in the targeted gene in hatched larvae. One promoter, Chom U6_b, showed the highest activity. These U6 gene promoters could be used to build CRISPR/Cas9-based genetic systems for the control of C. hominivorax. © 2023 The Authors. Insect Molecular Biology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.  aCRISPR/Cas9  aGenome editing  aPartnership for the goals - Goal 17  aPest control  aPLATAFORMA DE INVESTIGACIÓN EN SALUD ANIMAL - INIA  aScrewworm  aSustainable Development Goals (SDGs)  aU6 promoter1 aBASIKA, T.1 aWILLIAMSON, M. E.1 aFRESIA, P.1 aMENCHACA, A.1 aSCOTT, M. J.  tInsect Molecular Biology, 2023. Early View. https://doi.org/10.1111/imb.12875   -- OPEN ACCESS.