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

• Martina Crispo ^[1] ; Geraldine Schlapp ^[1] ; Magdalena Cárdenas-Rodríguez ^[1] ; Dolores González-Maciel ^[2] ; Martín Rumbo ^[2]
  1. [1] Institut Pasteur de Montevideo

     Institut Pasteur de Montevideo

     Uruguay

     
  2. [2] Universidad Nacional de La Plata

     Universidad Nacional de La Plata

     Argentina

     
• Localización: Electronic Journal of Biotechnology, ISSN-e 0717-3458, Vol. 16, Nº. 6, 2013, págs. 14-14
• Idioma: inglés
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• Resumen

  • 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.