Kretov Dmitry A, Shafik Andrew M, Cifuentes Daniel
Department of Biochemistry, Boston University School of Medicine, 72 East Concord Street, Boston, MA, 02118, USA.
Methods Mol Biol. 2018;1680:179-190. doi: 10.1007/978-1-4939-7339-2_12.
The ability to microinject small RNAs and mRNAs into zebrafish embryos, of different genetic backgrounds, allows for the precise dissection of microRNA processing pathways at the molecular level, while simultaneously provides insight into their physiologic role. Here, we apply such an approach to determine the impact of Argonaute 2 in the processing of miR-451, a vertebrate-specific microRNA required for terminal erythrocyte differentiation. This was achieved using fluorescent microRNA reporter sensor assays and phenotype rescue experiments.
将小RNA和mRNA显微注射到具有不同遗传背景的斑马鱼胚胎中的能力，使得在分子水平上精确剖析微小RNA加工途径成为可能，同时还能深入了解它们的生理作用。在此，我们应用这种方法来确定AGO2在miR-451加工过程中的作用，miR-451是终末红细胞分化所需的脊椎动物特异性微小RNA。这是通过荧光微小RNA报告传感器检测和表型拯救实验实现的。
