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. 2006 Apr 28;2(4):e65. doi: 10.1371/journal.pgen.0020065

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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Cells containing cassette 234-β-EGFP at RL5 were transfected with the Cre recombinase, and inverted sub-clones were identified using primer pairs p1/p2 (specific for the P orientation) and p1/p3 (specific for the N orientation).

(A) Location of the primers used to identify the inverted clones.

(B) P to N inversion: The histogram on the left illustrates expression profile of a clone with insertion in the P orientation before inversion. The histogram on the right illustrates expression 8 wk post-inversion. The ethidium bromide stain illustrates the PCR analysis demonstrating the inversion.

(C) Same as (B) but for the N to P inversion. This analysis clearly demonstrates that Cre recombination per se does not lead to silencing (see text). As reported previously, levels of expression in the N orientation at RL5 are lower than in the P orientation [32].

N, non-permissive orientation; P, permissive orientation.

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