Department of Developmental Biochemistry, Göttingen Center for Molecular Biosciences, Justus-von-Liebig-Weg 11, Göttingen, Germany.
Methods. 2010 May;51(1):146-51. doi: 10.1016/j.ymeth.2010.02.016. Epub 2010 Feb 21.
Localized mRNAs have been identified in a large variety of cell types where they contribute to the establishment of cell asymmetries and can function as cell fate determinants. In Xenopus, RNAs that localize to the vegetal cortex during oogenesis function in early embryonic patterning as well as in the development of primordial germ cells. Based on their temporal and spatial localization patterns, vegetally localizing RNAs are referred to as either early-pathway RNAs which transiently localize in the mitochondrial cloud, or as late-pathway RNAs. Vegetal RNA-localization is driven by cis-acting signal sequences that, in most cases, were found to reside in the 3'-UTRs and which are recognized by trans-acting localization factors. Here we describe the methods of how vegetal RNA-localization elements can be identified by injection of fluorescently-labeled or tagged RNAs.
已在多种细胞类型中鉴定出定位的 mRNAs,它们有助于建立细胞不对称性,并可作为细胞命运决定因素发挥作用。在非洲爪蟾中,卵母细胞发生过程中定位于植物皮质的 RNA 可在早期胚胎模式形成以及原始生殖细胞的发育中发挥作用。根据它们的时空定位模式,定位于植物的 RNA 被称为早期途径 RNA,它们在短暂时间内定位于线粒体云中,或者称为晚期途径 RNA。植物 RNA 的定位是由顺式作用信号序列驱动的,在大多数情况下,这些信号序列位于 3'UTR 中,并被反式作用定位因子识别。在这里,我们描述了通过注射荧光标记或标记 RNA 来鉴定植物 RNA 定位元件的方法。