Protocol to measure cleavage efficiency of the meiotic cohesin subunit Rec8 by separase in mouse oocytes using a biosensor

使用生物传感器测量小鼠卵母细胞中分离酶对减数分裂黏连蛋白亚基 Rec8 的裂解效率的方案

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作者:Elvira Nikalayevich, Katja Wassmann

Abstract

Here, we describe a biosensor to assess meiotic cohesin subunit Rec8 cleavage in mouse oocytes. We detail oocyte collection and microinjection of the mRNA expressing the biosensor. The biosensor is targeted to chromosomes and consists of two fluorophores flanking a Rec8 fragment containing separase cleavage sites. Cleavage leads to dissociation of one fluorophore from chromosomes, and the efficiency can be estimated by live imaging. We detail the use of this biosensor in mouse oocytes with or without Aurora B/C inhibitor. For complete details on the use and execution of this protocol, please refer to Nikalayevich et al. (2022).

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