Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line

Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to opti...

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Autores principales:	Crispo,Martina, Schlapp,Geraldine, Cárdenas-Rodríguez,Magdalena, González-Maciel,Dolores, Rumbo,Martín
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2013
Materias:	DNA concentration microinjection reporter systems
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600014
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spelling	oai:scielo:S0717-345820130006000142014-01-16Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice lineCrispo,MartinaSchlapp,GeraldineCárdenas-Rodríguez,MagdalenaGonzález-Maciel,DoloresRumbo,Martín DNA concentration microinjection reporter systems Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.16 n.6 20132013-11-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600014en10.2225/vol16-issue6-fulltext-3
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	DNA concentration microinjection reporter systems
spellingShingle	DNA concentration microinjection reporter systems Crispo,Martina Schlapp,Geraldine Cárdenas-Rodríguez,Magdalena González-Maciel,Dolores Rumbo,Martín Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
description	Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.
author	Crispo,Martina Schlapp,Geraldine Cárdenas-Rodríguez,Magdalena González-Maciel,Dolores Rumbo,Martín
author_facet	Crispo,Martina Schlapp,Geraldine Cárdenas-Rodríguez,Magdalena González-Maciel,Dolores Rumbo,Martín
author_sort	Crispo,Martina
title	Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_short	Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_full	Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_fullStr	Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_full_unstemmed	Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_sort	optimization of transgenesis conditions for the generation of cxcl2-luciferase reporter mice line
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2013
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600014
work_keys_str_mv	AT crispomartina optimizationoftransgenesisconditionsforthegenerationofcxcl2luciferasereportermiceline AT schlappgeraldine optimizationoftransgenesisconditionsforthegenerationofcxcl2luciferasereportermiceline AT cardenasrodriguezmagdalena optimizationoftransgenesisconditionsforthegenerationofcxcl2luciferasereportermiceline AT gonzalezmacieldolores optimizationoftransgenesisconditionsforthegenerationofcxcl2luciferasereportermiceline AT rumbomartin optimizationoftransgenesisconditionsforthegenerationofcxcl2luciferasereportermiceline
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Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line

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