Protocol to measure cleavage efficiency of the meiotic cohesin subunit Rec8 by separase in mouse oocytes using a biosensor - Archive ouverte HAL Access content directly
Journal Articles STAR Protocols Year : 2022

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

Abstract

Here we describe a biosensor to assess meiotic cohesin cleavage by live imaging in mouse oocytes. The biosensor is targeted to chromosomes and consists of two fluorophores flanking a fragment of Rec8 containing Separase cleavage sites. Cleavage of Rec8 leads to dissociation of one of the fluorophores from chromosomes which is used to estimate cleavage efficiency. We detail the use of this biosensor in mouse oocytes completing the first meiotic division in presence or absence of AuroraB/C inhibitor. For details on the application of this protocol, please refer to (Nikalayevich et al., 2022).
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Dates and versions

hal-03809570 , version 1 (12-10-2022)

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Elvira Nikalayevich, Katja Wassmann. Protocol to measure cleavage efficiency of the meiotic cohesin subunit Rec8 by separase in mouse oocytes using a biosensor. STAR Protocols, In press, 3 (4), pp.101714. ⟨10.1016/j.xpro.2022.101714⟩. ⟨hal-03809570⟩
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