The Mary Lyon Centre, MRC Harwell, Didcot, Oxon, UK.
Methods Mol Biol. 2023;2631:103-134. doi: 10.1007/978-1-0716-2990-1_4.
Targeted nucleases allow the production of many types of genetic mutations directly in the early embryo. However, the outcome of their activity is a repair event of unpredictable nature, and the founder animals that are produced are generally of a mosaic nature. Here, we present the molecular assays and genotyping strategies that will support the screening of the first generation for potential founders and the validation of positive animals in the subsequent generation, according to the type of mutation generated.
靶向核酸酶允许在早期胚胎中直接产生多种类型的遗传突变。然而,它们的活性的结果是一种不可预测的性质的修复事件,并且产生的创始动物通常是嵌合体性质的。在这里,我们提出了分子检测和基因分型策略,这些策略将根据产生的突变类型,支持第一代潜在创始动物的筛选和随后一代阳性动物的验证。