| Autor: |
D'Orazio FM; Institute of Cancer and Genomics Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK.; MRC London Institute of Medical Sciences and Faculty of Medicine, Imperial College, London, UK.; Institute of Clinical Sciences, Faculty of Medicine, Imperial College, London, UK., Jasiulewicz A; Institute of Cancer and Genomics Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK., Hadzhiev Y; Institute of Cancer and Genomics Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK., Müller F; Institute of Cancer and Genomics Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK. F.Mueller@bham.ac.uk. |
| Abstrakt: |
Here, we describe a fast and straightforward methodology to in vivo detect transcriptional activity in the early zebrafish germ line. We report how fluorescently labeled morpholinos, targeted to nascent early transcripts, can be used to track the onset of transcriptional events during early embryogenesis. This method could be applied to any tagged cell line in a developing early zebrafish embryo as long as the gene of interest is expressed at high enough level for morpholino detection and is expressed at the first and main wave of genome activation, for which the protocol has been verified. The protocol, in combination with genetic manipulation, allows studies of mechanisms driving zygotic genome activation (ZGA) in individual cells. The reported procedures apply to a broad range of purposes for zebrafish embryo manipulation in view of imaging nuclear molecules in specific cell types. |
