School of Biological Sciences, University of Adelaide, Adelaide, SA, Australia.
South Australian Research and Development Institute, Adelaide, SA, Australia.
Methods Mol Biol. 2022;2495:151-171. doi: 10.1007/978-1-0716-2301-5_9.
Sterile Insect Technique (SIT) is a biocontrol strategy that has been widely utilized to suppress or eradicate outbreak populations of insect pests such as tephritid fruit flies. As SIT is highly favored due to it being species-specific and environmentally friendly, there are constant efforts to improve the efficiency and efficacy of this method in particular at low pest densities; one of which is the use of genetically enhanced strains. Development of these desirable strains has been facilitated by the emergence of the CRISPR/Cas genome-editing technology that enables the rapid and precise genomic modification of non-model organisms. Here, we describe the manual microinjection of CRISPR/Cas9 reagents into tephritid pest Bactrocera tryoni (Queensland fruit fly) embryos to introduce ideal traits as well as the molecular methods used to detect successful mutagenesis.
sterile insect technique (sit) 是一种已被广泛用于抑制或根除害虫种群(如桔小实蝇)的生物防治策略。由于 sit 具有物种特异性和环境友好性,因此一直以来人们都在努力提高这种方法的效率和效果,特别是在害虫密度较低的情况下;其中一种方法是使用遗传增强株系。crispr/cas 基因组编辑技术的出现促进了这些理想株系的发展,该技术能够快速、精确地对非模式生物进行基因组修饰。在这里,我们描述了将 crispr/cas9 试剂手动微注射到桔小实蝇(昆士兰果蝇)胚胎中以引入理想性状的方法,以及用于检测成功诱变的分子方法。