Protocol for differential multi-omic analyses of distinct cell types in the mouse cerebral cortex

Here, we present a protocol for differential multi-omic analyses of distinct cell types in the developing mouse cerebral cortex. We describe steps for in utero electroporation, subsequent flow-cytometry-based isolation of developing mouse cortical cells, bulk RNA sequencing or quantitative liquid ch...

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Veröffentlicht in:STAR protocols 2024-03, Vol.5 (1), p.102793-102793, Article 102793
Hauptverfasser: Meka, Durga Praveen, Richter, Melanie, Rücker, Tabitha, Voss, Hannah, Rissiek, Anne, Krisp, Christoph, Kumar, Nisha Hemandhar, Schwanke, Birgit, Fornasiero, Eugenio F., Schlüter, Hartmut, Calderon de Anda, Froylan
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Sprache:eng
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Zusammenfassung:Here, we present a protocol for differential multi-omic analyses of distinct cell types in the developing mouse cerebral cortex. We describe steps for in utero electroporation, subsequent flow-cytometry-based isolation of developing mouse cortical cells, bulk RNA sequencing or quantitative liquid chromatography-tandem mass spectrometry, and bioinformatic analyses. This protocol can be applied to compare the proteomes and transcriptomes of developing mouse cortical cell populations after various manipulations (e.g., epigenetic). For complete details on the use and execution of this protocol, please refer to Meka et al. (2022).1 [Display omitted] •Labeling and isolation of mouse cortical cells by IUE and FACS, respectively•Simultaneous analysis of the cell population across different modalities in bulk•Comparison of cells from mouse embryos with genetic or pharmacological modification•Quantitative comparison of proteomes using LC-MS/MS and bioinformatics Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for differential multi-omic analyses of distinct cell types in the developing mouse cerebral cortex. We describe steps for in utero electroporation, subsequent flow-cytometry-based isolation of developing mouse cortical cells, bulk RNA sequencing or quantitative liquid chromatography-tandem mass spectrometry, and bioinformatic analyses. This protocol can be applied to compare the proteomes and transcriptomes of developing mouse cortical cell populations after various manipulations (e.g., epigenetic).
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2023.102793