CMMB 403 Lecture Notes - Lecture 30: Cre Recombinase, Microinjection, Knockout Mouse
6 Jan 2019
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A loxp site is a specific 34 base pair dna sequence. Chance that this sequence will occur naturally in the genome is very small. Naturally exists in bacteria that have circular dna. The cre-recombinase is an enzyme from a bacterial phage that recognizes two loxp sites and deletes the dna between them. Net effect: circular dna that is deleted. Cre enzyme comes from bacteriophage, and exists as a dimer. Cre and loxp each exists in different mice. Transgenic mice expressing cre recombinase in the tissue of interest. Mice with loxp sites on each side of the gene or region to be deleted. The pre transgenic mouse has a tissue-specific promoter driving cre is not targeted. You need to get these together in a single mouse, so both copies of the gene is floxed and the cre is in a single mouse or embryo that is your conditional mutant. To get the floxed gene homozygous together with the.
