Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method

Background: Production of transgenic animals is still a low-efficiency biotechnology, and simple alternatives should be used to improve the rate of transgenic bovine production by nuclear transfer. One such alternative is selecting the appropriate donor cell type and transfection method. Objective:...

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Autores principales:	Alinne G. Curcio, Fabiana F. Bressan, Carla S. Paes De Carvalho, Célia R. Quirino, Flavio V. Meirelles, Angelo J. B. Dias
Formato:	article
Lenguaje:	EN
Publicado:	Universidad de Antioquia 2019
Materias:	cloning epigenetics lipofection lentiviral transduction nuclear reprogramming Animal culture SF1-1100
Acceso en línea:	https://doaj.org/article/1da6cdeade074b7eafdc13f9dd554c23
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spelling	oai:doaj.org-article:1da6cdeade074b7eafdc13f9dd554c232021-12-01T19:15:23ZEfficiency of transgene expression in bovine cells varies according to cell type and gene transfer method2256-295810.17533/udea.rccp.v32n1a04https://doaj.org/article/1da6cdeade074b7eafdc13f9dd554c232019-03-01T00:00:00Zhttps://revistas.udea.edu.co/index.php/rccp/article/view/333061https://doaj.org/toc/2256-2958Background: Production of transgenic animals is still a low-efficiency biotechnology, and simple alternatives should be used to improve the rate of transgenic bovine production by nuclear transfer. One such alternative is selecting the appropriate donor cell type and transfection method. Objective: To investigate the effect of cell type (fetal or adult fibroblasts, and cumulus cells), and gene transfer method (lipofection and lentiviral transduction) on the incorporation, expression, and fluorescence intensity of transgene on bovine cells analyzed by flow cytometry. Methods: Fetal fibroblasts (FF), adult fibroblasts (AF), and cumulus cells (CC) were transfected using lipofection, or transduced using lentiviral particles produced with Green Fluorescent Protein (GFP) expressing plasmids, and analyzed by flow cytometry. Results: Lentiviral transduction resulted in higher transgene expression rates for all cell types (FF: 88.8 ± 0.98; AF: 91.6 ± 2.96; CC: 60.7% ± 14.7) compared to lipofection (FF: 17.8 ± 2.82; AF: 10.66 ± 0.65; CC: 3.9% ± 1.97). Cumulus cells showed lower transgene expression rates than the other cell types. Regarding fluorescence intensity, there was no significant difference between lipofection and lentiviral transduction; in both treatments, higher fluorescence intensity was obtained when adult cells were used instead of fetal cells. Conclusion: Gene transfer efficiency varies according to cell type, and gene transfer method, with lentiviral transduction achieving higher transgene expression rate, and adult fibroblasts showing better transgene expression.Alinne G. CurcioFabiana F. BressanCarla S. Paes De CarvalhoCélia R. QuirinoFlavio V. MeirellesAngelo J. B. DiasUniversidad de Antioquiaarticlecloningepigeneticslipofectionlentiviral transductionnuclear reprogrammingAnimal cultureSF1-1100ENRevista Colombiana de Ciencias Pecuarias, Vol 32, Iss 1, Pp 34-42 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	cloning epigenetics lipofection lentiviral transduction nuclear reprogramming Animal culture SF1-1100
spellingShingle	cloning epigenetics lipofection lentiviral transduction nuclear reprogramming Animal culture SF1-1100 Alinne G. Curcio Fabiana F. Bressan Carla S. Paes De Carvalho Célia R. Quirino Flavio V. Meirelles Angelo J. B. Dias Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
description	Background: Production of transgenic animals is still a low-efficiency biotechnology, and simple alternatives should be used to improve the rate of transgenic bovine production by nuclear transfer. One such alternative is selecting the appropriate donor cell type and transfection method. Objective: To investigate the effect of cell type (fetal or adult fibroblasts, and cumulus cells), and gene transfer method (lipofection and lentiviral transduction) on the incorporation, expression, and fluorescence intensity of transgene on bovine cells analyzed by flow cytometry. Methods: Fetal fibroblasts (FF), adult fibroblasts (AF), and cumulus cells (CC) were transfected using lipofection, or transduced using lentiviral particles produced with Green Fluorescent Protein (GFP) expressing plasmids, and analyzed by flow cytometry. Results: Lentiviral transduction resulted in higher transgene expression rates for all cell types (FF: 88.8 ± 0.98; AF: 91.6 ± 2.96; CC: 60.7% ± 14.7) compared to lipofection (FF: 17.8 ± 2.82; AF: 10.66 ± 0.65; CC: 3.9% ± 1.97). Cumulus cells showed lower transgene expression rates than the other cell types. Regarding fluorescence intensity, there was no significant difference between lipofection and lentiviral transduction; in both treatments, higher fluorescence intensity was obtained when adult cells were used instead of fetal cells. Conclusion: Gene transfer efficiency varies according to cell type, and gene transfer method, with lentiviral transduction achieving higher transgene expression rate, and adult fibroblasts showing better transgene expression.
format	article
author	Alinne G. Curcio Fabiana F. Bressan Carla S. Paes De Carvalho Célia R. Quirino Flavio V. Meirelles Angelo J. B. Dias
author_facet	Alinne G. Curcio Fabiana F. Bressan Carla S. Paes De Carvalho Célia R. Quirino Flavio V. Meirelles Angelo J. B. Dias
author_sort	Alinne G. Curcio
title	Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
title_short	Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
title_full	Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
title_fullStr	Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
title_full_unstemmed	Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
title_sort	efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method
publisher	Universidad de Antioquia
publishDate	2019
url	https://doaj.org/article/1da6cdeade074b7eafdc13f9dd554c23
work_keys_str_mv	AT alinnegcurcio efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod AT fabianafbressan efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod AT carlaspaesdecarvalho efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod AT celiarquirino efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod AT flaviovmeirelles efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod AT angelojbdias efficiencyoftransgeneexpressioninbovinecellsvariesaccordingtocelltypeandgenetransfermethod
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Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method

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