Recombinase Conditional

Key words RNAi, Conditional, ShRNA, Cre recombinase, TetR/O, Transgenic mice, Rosa26,... 

Delete the lox-STOP-lox cassette in vitro by incubating 1 pg conditional shRNA plasmid DNA with 4 units of Cre recom-binase (New England Biolabs) in a total volume of 50 pi lx Cre buffer at 37°C for 30 min. Inactivate the recombinase for 10 min at 70°C. Eliminate nonrecombined plasmids by EcoRI digestion (see Note 8). 

Zhang, Z. and Lutz, B. (2002) Cre recombinase-mediated inversion using lox66 and lox71 method to introduce conditional point mutations into the CREB-binding protein. Nucl. Acids Res. 30, e90. 

Le Y, Sauer B 2000 Conditional gene knockout using ere recombinase. Methods Mol Biol 136 477-485... 

P. Chambon, Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase, Proc. Natl. Acad. Sci. U.S.A. 1995, 92, 6991-6995. 

Similarly, in mice bearing a conditionally targeted vhl allele, Cre-recombinase-mediated excision results in activation of the HIF pathway in hepatocytes (47). In a further example, some sublines of Chinese hamster ovary-Kl cells selected for activating mutations in the HIF pathway were found to have vhl mutations, and expression of a wild-type human VHL gene restored HIF regulation to normal (48). 

---