Development of CRISPR/Cas9 System Delivery Methods and Their Applications in Insect Research
Since the emergence of the CRISPR/Cas9 gene editing system,there has been significant ad-vancement in understanding biological gene functions.CRISPR/Cas9 is typically delivered via microinjection into insect embryos for gene editing.However,due to diverse reproductive modes in insects,it is difficult to use traditional CRISPR/Cas9 delivery methods to study the gene functions of some insect species,such as cockroaches with rigid egg cases and viviparous aphids.In the current CRISPR/Cas9 technology,alternative delivery methods have been developed,which do not require injection into early-stage insect embryos and have been reported as ReMOT(receptor-mediated ovary transduction)and DIPA-CRISPR("direct parental"CRISPR).These methods allow the direct injection of editing reagents into the hemocoel of adult female in-dividuals,introducing genetic mutations effectively into developing oocytes and simplifying the gene editing process.They have been successfully applied to the study of insects such as Aedes aegypti,Blattella ger-manica,and Tribolium castaneum.This article reviews the structure,mechanism,and traditional delivery methods of the CRISPR/Cas9 system,as well as the application and prospects of the newly developed Re-MOT and DIPA-CRISPR delivery methods in insect research,aiming to provide guidance for gene editing in more species.
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