Počet záznamů: 1  

Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling. A Novel Polysome Fractionation Method

  1. 1.
    SYSNO ASEP0523873
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    Poddruh JČlánek ve WOS
    NázevIdentifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling. A Novel Polysome Fractionation Method
    Tvůrce(i) Mašek, T. (CZ)
    del Llano, Edgar (UZFG-Y) ORCID
    Gahurová, Lenka (UZFG-Y) ORCID
    Kubelka, Michal (UZFG-Y) RID, ORCID
    Šušor, Andrej (UZFG-Y) RID, ORCID
    Roučová, K. (CZ)
    Lin, C. J. (GB)
    Bruce, A. W. (CZ)
    Pospíšek, M. (CZ)
    Číslo článku1254
    Zdroj.dok.International Journal of Molecular Sciences. - : MDPI
    Roč. 21, č. 4 (2020)
    Poč.str.23 s.
    Forma vydáníOnline - E
    Jazyk dok.eng - angličtina
    Země vyd.CH - Švýcarsko
    Klíč. slovapolysome profiling ; polysome fractionation ; translatome ; mouse oocyte ; mouse zygote
    Vědní obor RIVEB - Genetika a molekulární biologie
    Obor OECDDevelopmental biology
    CEPGA19-13491S GA ČR - Grantová agentura ČR
    Způsob publikováníOpen access
    Institucionální podporaUZFG-Y - RVO:67985904
    UT WOS000522524400081
    EID SCOPUS85079586112
    DOI10.3390/ijms21041254
    AnotaceMeiotic maturation of oocyte relies on pre-synthesised maternal mRNA, the translation of which is highly coordinated in space and time. Here, we provide a detailed polysome profiling protocol that demonstrates a combination of the sucrose gradient ultracentrifugation in small SW55Ti tubes with the qRT-PCR-based quantification of 18S and 28S rRNAs in fractionated polysome profile. This newly optimised method, named Scarce Sample Polysome Profiling (SSP-profiling), is suitable for both scarce and conventional sample sizes and is compatible with downstream RNA-seq to identify polysome associated transcripts. Utilising SSP-profiling we have assayed the translatome of mouse oocytes at the onset of nuclear envelope breakdown (NEBD)-a developmental point, the study of which is important for furthering our understanding of the molecular mechanisms leading to oocyte aneuploidy. Our analyses identified 1847 transcripts with moderate to strong polysome occupancy, including abundantly represented mRNAs encoding mitochondrial and ribosomal proteins, proteasomal components, glycolytic and amino acids synthetic enzymes, proteins involved in cytoskeleton organization plus RNA-binding and translation initiation factors. In addition to transcripts encoding known players of meiotic progression, we also identified several mRNAs encoding proteins of unknown function. Polysome profiles generated using SSP-profiling were more than comparable to those developed using existing conventional approaches, being demonstrably superior in their resolution, reproducibility, versatility, speed of derivation and downstream protocol applicability.
    PracovištěÚstav živočišné fyziologie a genetiky
    KontaktJana Zásmětová, knihovna@iapg.cas.cz, Tel.: 315 639 554
    Rok sběru2021
    Elektronická adresahttps://www.mdpi.com/1422-0067/21/4/1254
Počet záznamů: 1  

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