Establishment of nestin transgenic mice and nestin expression in the organs

Objective To develop a nestin transgenic mice and study the distribution of nestin expression in the organs.Methods The three segments of nestin in full-length cDNA was amplified using human glioma cell line U251 cDNA as the template and cloned into the vector pBROAD3 containing a strong promoter ROSA26.The constructed vector,after identification with restriction enzyme and sequencing and removal of the prokaryotic sequences,were purified and injected into the fertilized eggs of mice.Transgenic mice were identified by PCR and the founder was maintained.Western blot analysis was used to detect the expression of nestin of the F3 transgenic mice and the wild-type ones in the vital organs(heart,lung,brain and kidney).Results The Nestin transgenic vector controlled by ROSA26 promoter was successfully constructed and validated by sequencing.Among the 34 newborn mice,2 founders were tested to be nestin-positive by PCR.Westem blot analysis showed that the F3 transgenic mice expressed high levels of nestin in the brain and lungs.Conclusion Nestin transgenic mice have been successfully established with stable nestin expression in the brain and lungs of the offspring mice,which can be useful for studying the functions of nestin in tumor metastasis,stemness maintenance and differentiation of cells.