| Registro completo |
| Provedor de dados: |
Electron. J. Biotechnol.
|
| País: |
Chile
|
| Título: |
Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
|
| Autores: |
Crispo,Martina
Schlapp,Geraldine
Cárdenas-Rodríguez,Magdalena
González-Maciel,Dolores
Rumbo,Martín
|
| Data: |
2013-11-01
|
| Ano: |
2013
|
| Palavras-chave: |
DNA concentration
Microinjection
Reporter systems
|
| Resumo: |
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.
|
| Tipo: |
Journal article
|
| Idioma: |
Inglês
|
| Identificador: |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600014
|
| Editor: |
Pontificia Universidad Católica de Valparaíso
|
| Formato: |
text/html
|
| Fonte: |
Electronic Journal of Biotechnology v.16 n.6 2013
|
|
