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De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements

Mosaic analysis by dual recombinase-mediated cassette exchange (MADR) is a technology that allows stable and locus-specific integration of transgenic elements into recipient cells carrying loxP and FRT sites. Nevertheless, most cell lines lack these recombination-specific sites. This protocol descri...

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Detalles Bibliográficos
Autores principales: Ayala-Sarmiento, Alberto E., Kobritz, Naomi, Breunig, Joshua J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757409/
https://www.ncbi.nlm.nih.gov/pubmed/33377078
http://dx.doi.org/10.1016/j.xpro.2020.100184
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author Ayala-Sarmiento, Alberto E.
Kobritz, Naomi
Breunig, Joshua J.
author_facet Ayala-Sarmiento, Alberto E.
Kobritz, Naomi
Breunig, Joshua J.
author_sort Ayala-Sarmiento, Alberto E.
collection PubMed
description Mosaic analysis by dual recombinase-mediated cassette exchange (MADR) is a technology that allows stable and locus-specific integration of transgenic elements into recipient cells carrying loxP and FRT sites. Nevertheless, most cell lines lack these recombination-specific sites. This protocol describes a method to introduce the minimum requirements into cells, leading to the generation of de novo primary MADR recipient cells or MADR “Proxy” cells. These cell lines allow the combinatorial use of a wide range of transgenic elements through MADR. For complete details on the use and execution of this protocol, please refer to Kim et al. (2019).
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spelling pubmed-77574092020-12-28 De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements Ayala-Sarmiento, Alberto E. Kobritz, Naomi Breunig, Joshua J. STAR Protoc Protocol Mosaic analysis by dual recombinase-mediated cassette exchange (MADR) is a technology that allows stable and locus-specific integration of transgenic elements into recipient cells carrying loxP and FRT sites. Nevertheless, most cell lines lack these recombination-specific sites. This protocol describes a method to introduce the minimum requirements into cells, leading to the generation of de novo primary MADR recipient cells or MADR “Proxy” cells. These cell lines allow the combinatorial use of a wide range of transgenic elements through MADR. For complete details on the use and execution of this protocol, please refer to Kim et al. (2019). Elsevier 2020-11-25 /pmc/articles/PMC7757409/ /pubmed/33377078 http://dx.doi.org/10.1016/j.xpro.2020.100184 Text en © 2020 The Authors 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
Ayala-Sarmiento, Alberto E.
Kobritz, Naomi
Breunig, Joshua J.
De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title_full De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title_fullStr De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title_full_unstemmed De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title_short De Novo Generation of Murine and Human MADR Recipient Cell Lines for Locus-Specific, Stable Integration of Transgenic Elements
title_sort de novo generation of murine and human madr recipient cell lines for locus-specific, stable integration of transgenic elements
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757409/
https://www.ncbi.nlm.nih.gov/pubmed/33377078
http://dx.doi.org/10.1016/j.xpro.2020.100184
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