Revathi Kandan, Subramaniam Kuppuswamy
Department of Biotechnology, Indian Institute of Technology Madras, Chennai, Tamil Nadu, India.
MicroPubl Biol. 2022 Oct 20;2022. doi: 10.17912/micropub.biology.000647. eCollection 2022.
-mediated single-copy integration (MosSCI) in relies on the introduction of plasmid constructs into the germ line. Such plasmids form extrachromosomal arrays containing multiple copies of the transgene. Presently, one positive-selection and four negative-selection reporters are used to identify animals that carry the integrated transgene but not the array. Even with four reporters, the negative selection is inefficient. Here, we show that the expression of the toxic protein PEEL-1 from a transgene containing the endogenous introns kills all array-carrying animals, which facilitates efficient selection of animals carrying the integrated transgene.
在[具体物种]中，介导的单拷贝整合（MosSCI）依赖于将质粒构建体导入生殖系。此类质粒形成含有转基因多个拷贝的染色体外阵列。目前，一个阳性选择和四个阴性选择报告基因用于鉴定携带整合转基因但不携带阵列的动物。即便有四个报告基因，阴性选择效率依然低下。在此，我们表明，来自含有内源性内含子的转基因的毒性蛋白PEEL-1的表达会杀死所有携带阵列的动物，这有助于高效选择携带整合转基因的动物。
