McDaniel Stephen L, Harrison Melissa M
Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison WI 53706, USA.
Bio Protoc. 2019 Jul 5;9(13):e3296. doi: 10.21769/BioProtoc.3296.
The early embryo of exists as a rapidly dividing syncytium of nuclei that are transcriptionally silent. Maternally deposited factors are required to awaken the genome and assist in the transition from maternal to zygotic control of development. Because many of these essential factors are maternally deposited and the early nuclear divisions are so rapid, it has been difficult to assess the functional role of transcription factors at discrete points in early embryonic development. To address this issue, we have developed an optogenetic system that can rapidly and reversibly inactivate transcription factors with nuclear-cycle resolution. The temporal precision enabled by this technique will allow a mechanistic understanding of how transcription factors function together to control genome activation and patterning in the early embryo and is likely broadly applicable to factors throughout embryogenesis.
[物种名称]的早期胚胎以快速分裂的合胞体形式存在,其中的细胞核处于转录沉默状态。需要母源沉积因子来唤醒基因组,并协助从母体对发育的控制过渡到合子对发育的控制。由于许多这些必需因子是母源沉积的,并且早期核分裂非常迅速,因此很难评估转录因子在早期胚胎发育离散阶段的功能作用。为了解决这个问题,我们开发了一种光遗传学系统,该系统可以以核周期分辨率快速且可逆地使转录因子失活。这项技术所具备的时间精度将有助于从机制上理解转录因子如何共同作用以控制早期胚胎中的基因组激活和模式形成,并且可能广泛适用于整个胚胎发育过程中的各种因子。