Cre recombinase microinjection for single-cell tracing and localised gene targeting

Tracing and manipulating cells in embryos are essential to understand development. Lipophilic dye microinjections, viral transfection and iontophoresis have been key to map the origin of the progenitor cells that form the different organs in the postimplantation mouse embryo. These techniques requir...

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Detalles Bibliográficos
Autores: Sendra, Miquel, Hourcade, Juan de Dios, Temiño, Susana, Sarabia, Antonio J., Ocaña, Oscar H., Domı́nguez, Jorge N., Torres, Miguel
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2022
País:España
Institución:Universidad de Jaén
Repositorio:RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaén
OAI Identifier:oai:ruja.ujaen.es:10953/7298
Acceso en línea:https://hdl.handle.net/10953/7298
Access Level:acceso abierto
Palabra clave:Fate mapping, Clonal analysis, Labelling, Cre recombinase microinjection, Mouse
2402 Desarrollo embrionario
2406 Biología molecular
2407 Biología celular
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repository_id_str
spelling Cre recombinase microinjection for single-cell tracing and localised gene targetingSendra, MiquelHourcade, Juan de DiosTemiño, SusanaSarabia, Antonio J.Ocaña, Oscar H.Domı́nguez, Jorge N.Torres, MiguelFate mapping, Clonal analysis, Labelling, Cre recombinase microinjection, Mouse2402 Desarrollo embrionario2406 Biología molecular2407 Biología celularTracing and manipulating cells in embryos are essential to understand development. Lipophilic dye microinjections, viral transfection and iontophoresis have been key to map the origin of the progenitor cells that form the different organs in the postimplantation mouse embryo. These techniques require advanced manipulation skills and only iontophoresis, a demanding approach of limited efficiency, has been used for single-cell labelling. Here, we perform lineage tracing and local gene ablation using cell-permeant Cre recombinase (TAT-Cre) microinjection. First, we map the fate of undifferentiated progenitors to the different heart chambers. Then, we achieve single-cell recombination by titrating the dose of TAT-Cre, which allows clonal analysis of nascent mesoderm progenitors. Finally, injecting TAT-Cre to Mycnflox/flox embryos in the primitive heart tube revealed that Mycn plays a cell-autonomous role in maintaining cardiomyocyte proliferation. This tool will help researchers identify the cell progenitors and gene networks involved in organ development, helping to understand the origin of congenital defects.The Company of Biologists202620262022info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttps://hdl.handle.net/10953/7298reponame:RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaéninstname:Universidad de JaénInglésDevelopmentAttribution-NonCommercial-NoDerivs 3.0 Spainhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/info:eu-repo/semantics/openAccessoai:ruja.ujaen.es:10953/72982026-06-24T12:41:07Z
dc.title.none.fl_str_mv Cre recombinase microinjection for single-cell tracing and localised gene targeting
title Cre recombinase microinjection for single-cell tracing and localised gene targeting
spellingShingle Cre recombinase microinjection for single-cell tracing and localised gene targeting
Sendra, Miquel
Fate mapping, Clonal analysis, Labelling, Cre recombinase microinjection, Mouse
2402 Desarrollo embrionario
2406 Biología molecular
2407 Biología celular
title_short Cre recombinase microinjection for single-cell tracing and localised gene targeting
title_full Cre recombinase microinjection for single-cell tracing and localised gene targeting
title_fullStr Cre recombinase microinjection for single-cell tracing and localised gene targeting
title_full_unstemmed Cre recombinase microinjection for single-cell tracing and localised gene targeting
title_sort Cre recombinase microinjection for single-cell tracing and localised gene targeting
dc.creator.none.fl_str_mv Sendra, Miquel
Hourcade, Juan de Dios
Temiño, Susana
Sarabia, Antonio J.
Ocaña, Oscar H.
Domı́nguez, Jorge N.
Torres, Miguel
author Sendra, Miquel
author_facet Sendra, Miquel
Hourcade, Juan de Dios
Temiño, Susana
Sarabia, Antonio J.
Ocaña, Oscar H.
Domı́nguez, Jorge N.
Torres, Miguel
author_role author
author2 Hourcade, Juan de Dios
Temiño, Susana
Sarabia, Antonio J.
Ocaña, Oscar H.
Domı́nguez, Jorge N.
Torres, Miguel
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Fate mapping, Clonal analysis, Labelling, Cre recombinase microinjection, Mouse
2402 Desarrollo embrionario
2406 Biología molecular
2407 Biología celular
topic Fate mapping, Clonal analysis, Labelling, Cre recombinase microinjection, Mouse
2402 Desarrollo embrionario
2406 Biología molecular
2407 Biología celular
description Tracing and manipulating cells in embryos are essential to understand development. Lipophilic dye microinjections, viral transfection and iontophoresis have been key to map the origin of the progenitor cells that form the different organs in the postimplantation mouse embryo. These techniques require advanced manipulation skills and only iontophoresis, a demanding approach of limited efficiency, has been used for single-cell labelling. Here, we perform lineage tracing and local gene ablation using cell-permeant Cre recombinase (TAT-Cre) microinjection. First, we map the fate of undifferentiated progenitors to the different heart chambers. Then, we achieve single-cell recombination by titrating the dose of TAT-Cre, which allows clonal analysis of nascent mesoderm progenitors. Finally, injecting TAT-Cre to Mycnflox/flox embryos in the primitive heart tube revealed that Mycn plays a cell-autonomous role in maintaining cardiomyocyte proliferation. This tool will help researchers identify the cell progenitors and gene networks involved in organ development, helping to understand the origin of congenital defects.
publishDate 2022
dc.date.none.fl_str_mv 2022
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/10953/7298
url https://hdl.handle.net/10953/7298
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Development
dc.rights.none.fl_str_mv Attribution-NonCommercial-NoDerivs 3.0 Spain
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivs 3.0 Spain
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv The Company of Biologists
publisher.none.fl_str_mv The Company of Biologists
dc.source.none.fl_str_mv reponame:RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaén
instname:Universidad de Jaén
instname_str Universidad de Jaén
reponame_str RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaén
collection RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaén
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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