Mk. Ray et al., Development of a transgenic mouse model using rat insulin promoter to drive the expression of CRE recombinase in a tissue-specific manner, INT J PANCR, 25(3), 1999, pp. 157-163
Background: Tissue-specific ablation of a gene using the Cre-loxP system ha
s been used as an important tool to define its role, in addition to the tot
al ablation, to avoid the embryonic lethality in case of wide expression of
the target gene.
Methods: The RIP-Cre genetic construct was generated by standard subcloning
techniques and microinjected into one cell embryo to develop the transgeni
c mouse line. Transgenic mice were screened by polymerase chain reaction (P
CR) using DNA isolated from tell digestion. Tissue specificity of RIP was d
emonstrated by transient transfection of RIP-lacZ construct to NIT-1 cells
(mouse insulinoma cell line) in vitro.
Results: The 448 nucleotides of RIP were sufficient for beta-cell specific
expression of the reporter gene as evidenced by the presence of blue color
in the nucleus of NIT-1 cells. Isolated RIP-Cre transgene was microinjected
, and PCR screening identified two independent lines of transgenic mice. Ti
ssue specificity of RIP was demonstrated by reverse transcriptase polymeras
e chain reaction (RT-PCR) using the islet RNA from the transgenic mice.
Conclusion: We have established a tissue-specific transgenic mouse model us
ing Cre recombinase linked to rat insulin promoter (RIP) to drive the expre
ssion of the reporter gene specifically in the beta-cells. The RIP-Cre tran
sgenic mice will allow beta-cell specific ablation of target gene(s) to def
ine its role in the regulation of islet physiology.