RNA-Fluorescence In Situ Hybridization for the Detection of RNA in Mammalian Oocytes and Embryos

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985904%3A_____%2F25%3A00638242" target="_blank" >RIV/67985904:_____/25:00638242 - isvavai.cz</a>

Výsledek na webu

<a href="https://link.springer.com/protocol/10.1007/978-1-0716-4658-8_5" target="_blank" >https://link.springer.com/protocol/10.1007/978-1-0716-4658-8_5</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/978-1-0716-4658-8_5" target="_blank" >10.1007/978-1-0716-4658-8_5</a>

Jazyk výsledku

angličtina

Název v původním jazyce

RNA-Fluorescence In Situ Hybridization for the Detection of RNA in Mammalian Oocytes and Embryos

Popis výsledku v původním jazyce

RNA metabolism plays an essential role in the development of both oocytes and embryos. Their dependence on stored maternal transcripts is due to a long period of silenced transcription in which the oocyte undergoes meiotic maturation and fertilization. Although maternal RNAs are unusually stable, they should be replaced by “zygotic” transcripts during the transition from oocyte to zygote. Analysis of ncRNA and mRNA distribution in the oocyte can provide clues to the fate of RNA in the single-cell environment.nOur work focuses on the visualization of the subcellular distribution of specific RNAs in mammalian oocytes and early embryos. The localization of many RNAs in the oocyte and embryo is still not fully understood. In this chapter, we describe an optimized protocol for RNAscope, a type of RNA fluorescence in situ hybridization (RNA FISH), which is a valuable technique for exploring mammalian oocytes and embryos. The method is based on a specific probe design strategy that enables signal amplification together with simultaneous background suppression. Additionally, it is suitable for quantitative spatio-temporal analysis of specific RNA transcripts. RNAscope, a simple and a reliable protocol, contributes to advancing our understanding of RNA biology in the context of germ cell development.

Název v anglickém jazyce

RNA-Fluorescence In Situ Hybridization for the Detection of RNA in Mammalian Oocytes and Embryos

Popis výsledku anglicky

RNA metabolism plays an essential role in the development of both oocytes and embryos. Their dependence on stored maternal transcripts is due to a long period of silenced transcription in which the oocyte undergoes meiotic maturation and fertilization. Although maternal RNAs are unusually stable, they should be replaced by “zygotic” transcripts during the transition from oocyte to zygote. Analysis of ncRNA and mRNA distribution in the oocyte can provide clues to the fate of RNA in the single-cell environment.nOur work focuses on the visualization of the subcellular distribution of specific RNAs in mammalian oocytes and early embryos. The localization of many RNAs in the oocyte and embryo is still not fully understood. In this chapter, we describe an optimized protocol for RNAscope, a type of RNA fluorescence in situ hybridization (RNA FISH), which is a valuable technique for exploring mammalian oocytes and embryos. The method is based on a specific probe design strategy that enables signal amplification together with simultaneous background suppression. Additionally, it is suitable for quantitative spatio-temporal analysis of specific RNA transcripts. RNAscope, a simple and a reliable protocol, contributes to advancing our understanding of RNA biology in the context of germ cell development.

Druh

C - Kapitola v odborné knize

CEP obor

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OECD FORD obor

10605 - Developmental biology

Projekt

<a href="/cs/project/GA24-12217S" target="_blank" >GA24-12217S: Na tango jsou potřeba dva až tři: Genomické interakce a fenotypické vlastnosti u mezidruhových hybridů a polyploidů.</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název knihy nebo sborníku

Mammalian Oocyte Development. Methods and Protocols

ISBN

978-1-0716-4657-1

Počet stran výsledku

10

Strana od-do

57-66

Počet stran knihy

211

Název nakladatele

Humana Press

Místo vydání

New York

Kód UT WoS kapitoly

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