Abstract：
The Oriental fruit fly, Bactrocera dorsalis (Hendel), is a highly invasive pest of quarantine importance affecting the global fruit trade. In managing B. dorsalis, methods like cultural, biological, chemical, sterile insect technique (SIT), and semiochemical-mediated attract-and-kill are in use with varying success. The SIT approach is the method of choice for a chemical-free, long-term suppression of B. dorsalis, followed in many countries across the globe. The nonspecific mutations caused by irradiation affect the overall fitness of flies, thus requiring a more precise method for a heritable, fitness-not-compromising approach. In this regard, CRISPR/Cas9-mediated genome editing enables the creation of mutations at the precise genomic location/s through RNA-guided dsDNA cleavage. Of late, DNA-free editing employing ribonucleoprotein complex (RNP) is preferred to validate the target genes at G0 stage embryos in insects. It requires characterizing genomic edits from adults after completing their life cycle, which may entail a few days to months, depending on longevity. Additionally, edit characterization is required from each individual, as edits are unique. Therefore, all RNP-microinjected individuals must be maintained until the end of their life cycle, irrespective of editing. To overcome this impediment, we predetermine the genomic edits from the shed tissues, such as pupal cases, to maintain only edited individuals. In this study, we have shown the utility of pupal cases from five males and females of B. dorsalis to predetermine the genomic edits, which corroborated the edits from the respective adults.
摘要：
东方果蝇，dorsalis（Hendel），是一种高度侵入性的害虫，具有重要的检疫意义，影响着全球水果贸易。在管理背芽孢杆菌时，文化等方法，生物，化学,昆虫不育技术（SIT），和信息素介导的吸引和杀死正在使用，并取得了不同的成功。SIT方法是无化学品的选择方法，长期抑制背芽孢杆菌，在全球许多国家紧随其后。辐射引起的非特异性突变会影响果蝇的整体适应性，因此需要一种更精确的可遗传方法，健身不妥协的方法。在这方面,CRISPR/Cas9介导的基因组编辑能够通过RNA引导的dsDNA切割在精确的基因组位置产生突变。迟到了，优选采用核糖核蛋白复合物(RNP)的无DNA编辑来验证昆虫中G0期胚胎的靶基因。它需要在完成生命周期后对成年人的基因组编辑进行表征，这可能需要几天到几个月，取决于长寿。此外,每个人都需要编辑特征，因为编辑是独一无二的。因此,所有注射RNP的个体都必须维持到生命周期结束，不管编辑。为了克服这个障碍，我们从脱落的组织中预先确定基因组编辑，例如p箱，只保留编辑过的个人。在这项研究中,我们已经显示了从五个男性和女性的背芽孢杆菌的p病例的效用，以预先确定的基因组编辑，这证实了各自成年人的编辑。
