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

Martina Crispo, Geraldine Schlapp, Magdalena Cárdenas-Rodríguez, Dolores González-Maciel, Martín Rumbo
2013 Electronic Journal of Biotechnology  
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
more » ... 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. How to reference this article: CRISPO, M.; SCHLAPP, G.; CÁRDENAS-RODRIGUEZ, M.; GONZÁLEZ-MACIEL, D. and RUMBO, M. (2013). Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line. Electronic Journal of Biotechnology, vol. 16, no. 6. http://dx.
doi:10.2225/vol16-issue6-fulltext-3 fatcat:v2gh2c6e5rfv7ijny5yvczh23u