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Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy

Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to p...

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Detalles Bibliográficos
Autores principales: Laukoter, Susanne, Amberg, Nicole, Pauler, Florian M., Hippenmeyer, Simon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757670/
https://www.ncbi.nlm.nih.gov/pubmed/33377108
http://dx.doi.org/10.1016/j.xpro.2020.100215
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author Laukoter, Susanne
Amberg, Nicole
Pauler, Florian M.
Hippenmeyer, Simon
author_facet Laukoter, Susanne
Amberg, Nicole
Pauler, Florian M.
Hippenmeyer, Simon
author_sort Laukoter, Susanne
collection PubMed
description Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b).
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spelling pubmed-77576702020-12-28 Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy Laukoter, Susanne Amberg, Nicole Pauler, Florian M. Hippenmeyer, Simon STAR Protoc Protocol Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b). Elsevier 2020-12-16 /pmc/articles/PMC7757670/ /pubmed/33377108 http://dx.doi.org/10.1016/j.xpro.2020.100215 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Laukoter, Susanne
Amberg, Nicole
Pauler, Florian M.
Hippenmeyer, Simon
Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title_full Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title_fullStr Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title_full_unstemmed Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title_short Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
title_sort generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757670/
https://www.ncbi.nlm.nih.gov/pubmed/33377108
http://dx.doi.org/10.1016/j.xpro.2020.100215
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