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

microinjection
DNA concentration
reporter systems

How to Cite

1.
Crispo M, Schlapp G, Cárdenas-Rodríguez M, González-Maciel D, Rumbo M. Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line. Electron. J. Biotechnol. [Internet]. 2013 Nov. 15 [cited 2024 Dec. 11];16(6). Available from: https://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/v16n6-3

Abstract

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/ml group, 5 in 38 alive pups in the 1 ng/ml group and 1 in 21 alive pups in the 4 ng/ml group (p < 0.1). The overall transgenic efficiency was higher for the 1 ng/ml 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.

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