Pediatric Genomics Discovery Program, Department of Pediatrics and Genetics, Yale University School of Medicine, New Haven, Connecticut 06510, USA
Pediatric Genomics Discovery Program, Department of Pediatrics and Genetics, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Cold Spring Harb Protoc. 2022 Apr 1;2022(4):Pdb.prot107644. doi: 10.1101/pdb.prot107644.
Microinjection is an important technique used to study development in the oocyte and early embryo. In , substances such as DNA, mRNA, and morpholino oligonucleotides have traditionally been injected into , because of their large embryo size and the relatively long time from their fertilization to first division. In the past few decades, has become an important model in developmental biology; it is particularly useful in genetic studies. The advent and rapid development of CRISPR-Cas9 technology has provided an array of targeted gene manipulations for which is particularly suited. The equipment and protocol for microinjection is broadly transferable from There are important differences between the species to consider, however, including the smaller embryo size and faster embryo development time in There are a number of solutions and reagents that differ in concentration and composition as well. Here we describe a microinjection protocol specifically for studies in .
显微注射是研究卵母细胞和早期胚胎发育的重要技术。在这种情况下,由于胚胎较大,从受精到第一次分裂的时间相对较长,传统上向 中注射 DNA、mRNA 和吗啉代寡核苷酸等物质。在过去的几十年中, 已成为发育生物学中的重要模型;它在遗传研究中特别有用。CRISPR-Cas9 技术的出现和快速发展为 提供了一系列靶向基因操作,这是特别适合的。 显微注射的设备和方案可以广泛从 中转移过来,但是,需要考虑到这两个物种之间存在重要差异,包括 胚胎较小,胚胎发育时间较快。此外,在浓度和组成方面也有一些不同的解决方案和试剂。在这里,我们描述了一个专门用于 研究的显微注射方案。