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Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis

BACKGROUND: In ovo electroporation is a widely used technique to study gene function in developmental biology. Despite the widespread acceptance of this technique, no genome-wide analysis of the effects of in ovo electroporation, principally the current applied across the tissue and exogenous vector...

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Autores principales: Farley, Emma K, Gale, Emily, Chambers, David, Li, Meng
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105949/
https://www.ncbi.nlm.nih.gov/pubmed/21527010
http://dx.doi.org/10.1186/1749-8104-6-17
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author Farley, Emma K
Gale, Emily
Chambers, David
Li, Meng
author_facet Farley, Emma K
Gale, Emily
Chambers, David
Li, Meng
author_sort Farley, Emma K
collection PubMed
description BACKGROUND: In ovo electroporation is a widely used technique to study gene function in developmental biology. Despite the widespread acceptance of this technique, no genome-wide analysis of the effects of in ovo electroporation, principally the current applied across the tissue and exogenous vector DNA introduced, on endogenous gene expression has been undertaken. Here, the effects of electric current and expression of a GFP-containing construct, via electroporation into the midbrain of Hamburger-Hamilton stage 10 chicken embryos, are analysed by microarray. RESULTS: Both current alone and in combination with exogenous DNA expression have a small but reproducible effect on endogenous gene expression, changing the expression of the genes represented on the array by less than 0.1% (current) and less than 0.5% (current + DNA), respectively. The subset of genes regulated by electric current and exogenous DNA span a disparate set of cellular functions. However, no genes involved in the regional identity were affected. In sharp contrast to this, electroporation of a known transcription factor, Dmrt5, caused a much greater change in gene expression. CONCLUSIONS: These findings represent the first systematic genome-wide analysis of the effects of in ovo electroporation on gene expression during embryonic development. The analysis reveals that this process has minimal impact on the genetic basis of cell fate specification. Thus, the study demonstrates the validity of the in ovo electroporation technique to study gene function and expression during development. Furthermore, the data presented here can be used as a resource to refine the set of transcriptional responders in future in ovo electroporation studies of specific gene function.
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spelling pubmed-31059492011-06-02 Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis Farley, Emma K Gale, Emily Chambers, David Li, Meng Neural Dev Research Article BACKGROUND: In ovo electroporation is a widely used technique to study gene function in developmental biology. Despite the widespread acceptance of this technique, no genome-wide analysis of the effects of in ovo electroporation, principally the current applied across the tissue and exogenous vector DNA introduced, on endogenous gene expression has been undertaken. Here, the effects of electric current and expression of a GFP-containing construct, via electroporation into the midbrain of Hamburger-Hamilton stage 10 chicken embryos, are analysed by microarray. RESULTS: Both current alone and in combination with exogenous DNA expression have a small but reproducible effect on endogenous gene expression, changing the expression of the genes represented on the array by less than 0.1% (current) and less than 0.5% (current + DNA), respectively. The subset of genes regulated by electric current and exogenous DNA span a disparate set of cellular functions. However, no genes involved in the regional identity were affected. In sharp contrast to this, electroporation of a known transcription factor, Dmrt5, caused a much greater change in gene expression. CONCLUSIONS: These findings represent the first systematic genome-wide analysis of the effects of in ovo electroporation on gene expression during embryonic development. The analysis reveals that this process has minimal impact on the genetic basis of cell fate specification. Thus, the study demonstrates the validity of the in ovo electroporation technique to study gene function and expression during development. Furthermore, the data presented here can be used as a resource to refine the set of transcriptional responders in future in ovo electroporation studies of specific gene function. BioMed Central 2011-04-28 /pmc/articles/PMC3105949/ /pubmed/21527010 http://dx.doi.org/10.1186/1749-8104-6-17 Text en Copyright ©2011 Farley et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Farley, Emma K
Gale, Emily
Chambers, David
Li, Meng
Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title_full Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title_fullStr Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title_full_unstemmed Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title_short Effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
title_sort effects of in ovo electroporation on endogenous gene expression: genome-wide analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105949/
https://www.ncbi.nlm.nih.gov/pubmed/21527010
http://dx.doi.org/10.1186/1749-8104-6-17
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