Cas9/guide-RNA curbs "vector" mosquitoes from spreading viruses

When summer is approaching, people begin to fall into the fear of being dominated by mosquitoes.

Recently, "Nature Communications" magazine published an article entitled Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes.

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As we all know, gRNA and Cas9 nuclease are the right-hand arms of the CRISPR/Cas9 system for gene editing.

It can be seen that gRNA, as a type of non-coding RNA, is essential for gene editing and requires type III RNA polymerase to be synthesized by transcription.

Based on this, in order to evaluate the efficiency of the new CRISPR/Cas9 gene editing tool, the researchers performed functional verification of the "U6 promoter" that expresses Cas9 and gRNA in Culex mosquitoes in vitro and in vivo.

Validation of CRISPR reagent in Culex quinquefasciatus ovarian cell line

Of course, whether this "plasmid toolkit" can be a valuable tool against Culex mosquitoes needs further verification.

Phenotype, molecular and functional verification of vasa-Cas9 transgene

　　Subsequently, the researchers optimized the Cas9/guide-RNA expression "plasmid kit" by modifying the gRNA structural sequence, and successfully established a Cas9 gene transgenic line.

　　To this end, the researchers evaluated the effect of gRNA "Loop" scaffold variants on the transgenic efficiency of Drosophila.

　　GRNA scaffold variants improve gene drive efficiency in Drosophila melanogaster

　　In short, this breakthrough research has greatly revolutionized traditional gene editing technology.