Journal of Mosquito Research 2024, Vol.14, No.2, 76-86 http://emtoscipublisher.com/index.php/jmr 82 such as the Cas9/gRNA-mediated gene-drive rescue system developed for Anopheles stephensi, have shown promise in population modification and suppression. However, the long-term ecological impacts of such interventions are not fully understood, and there is a risk of disrupting local ecosystems and non-target species (Adolfi et al., 2020). Figure 3 Agcd gene, pCO37 gene-drive construct, and resulting phenotypes (Adopted from Carballar-Lejarazú et al., 2020) Image caption: (A) Agcd gene: maroon blocks, exons (E1-4); empty blocks, 3’and 5’-untranslated regions (UTR); thick black line, introns and intergenic DNA. pCO37 plasmid: maroon blocks, homology arms from the Agcd gene; blue blocks, dominant marker gene components (3XP3 and CFP); tan blocks, drive components (nanos promoter and SpCas9 protein-encoding sequences); green blocks, guide RNA components (U6 promoter and gRNA sequence); dark gray block, φC31 attP “docking” site. Genes and features of pCO37 are not to scale and approximate sizes of components in kilobases are listed in the Materials and Methods. Recombination resulting from HDR initiated at the SpCaS9/gRNA-mediated cut site (broken-line arrow) occurring within the pink-shaded regions results in integration of the gene-drive construct. (B) CFP+ (blue arrow) and homozygous Agcd-mutant (red arrow) phenotypes in larvae and (C) Agcd-mutant (red arrow) phenotype in pupae. (D) Homozygous Agcd mutant phenotype “red eye” (red arrows) in adults. Approximate image magnifications for B, C, and D are 20, 10, and 20×, respectively (Adopted from Carballar-Lejarazú et al., 2020)
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