Hi all just wondering if lox-p sites interfere with gene function?
Most of the time, loxp sites are inserted in introns around a critical exon. This should prevent interference with the orginal gene function. Once a mouse is made, you can test for the function of the gene product (Northern, PCR, or western) in the floxed/floxed cre neg animal.
Just one more for you I understand that the neo cassette can also alter gene function, causing a hypermorphic allele. is this true as well?
That is true
Look at the example papers below, in which the presence of a neo-cassette in the gene of SLC6A1, the neuronal GABA transporter 1, results in almost complete knockout of the transporter expression.
GABA transporter deficiency causes tremor, ataxia, nervousness, and increased GABA-induced tonic conductance in cerebellum.
Chiu CS, Brickley S, Jensen K, Southwell A, Mckinney S, Cull-Candy S, Mody I, Lester HA.
J Neurosci. 2005 Mar 23;25(12):3234-45.
GABA transporter-1 (GAT1)-deficient mice: differential tonic activation of GABAA versus GABAB receptors in the hippocampus.
Jensen K, Chiu CS, Sokolova I, Lester HA, Mody I.
J Neurophysiol. 2003 Oct;90(4):2690-701. Epub 2003 Jun 18.
Ok so if utilize a flipase cross to delete my frt-neo cassette then I can bypass these effects?
Sorry the reply took so long. The cross to a flipase mouse will remove the neocassette in the intron. Here are two examples, one from literature, one from Jackson lab.
Happy mouse breeding.