Strategie grantové aplikace
Základy vědecké práce
Pokroky v molekulární biologii
Předběžná náplň práce
Máme vybrané kandidátní geny, u kterých budeme sledovat expresi v průběhu meiotické maturace a ve včasném embryu. Také budeme studovat jejich roli pro fyziologii buňky.
Předběžná náplň práce v anglickém jazyce
We will identify potential players in oocyte an embryo development. In this aim, we plan to characterize phenotypic effect of the selected ncRNA. This makes knockdown and overexpression experiments crucial to understanding the roles of ncRNAs in vivo. To evaluate function in the mouse oocyte and early embryo development, the candidate transcript will be depleted by microinjection of dsRNA.As an alternative antisense DNA oligos will be used to induce RNase H activity in the nucleus (Vickers, 2003). To disrupt ncRNA function microinjection of the antisense oligos to the growing oocytes or zygotes targeting splice sites (Ulitsky, 2011) will be used. Inducing overexpression of the ncRNA by microinjection of in vitro transcribed RNA might show effect on the studied system. All microinjections will be with appropriate controls (e.g. targeting Egfp, Malat1 RNAs) and qPCR validation.