How does FRT Flippase work?
When there are two FRTs sequences in tandem, after inducing flippase the DNA sequence that is between these two sites will be deleted. If two FRT sites are facing each other (or looking away), the DNA that sits in between them can be inverted after induction of flippase.
What are Cre mice?
A Cre mouse contains a Cre recombinase transgene under the direction of a tissue-specific promoter; a loxP mouse contains two loxP sites that flank a genomic segment of interest, the “floxed” locus. Typically, Cre and loxP mice are produced by using transgenic technology (Nagy 2000).
Where is Flippase found?
the membrane
Flippases (rarely spelled flipases) are transmembrane lipid transporter proteins located in the membrane which belong to ABC transporter or P4-type ATPase families.
What kind of recombinase is used in FLP?
It involves using flippase (FLP) recombinase, derived from the yeast Saccharomyces cerevisiae (Sadowski 1995). FLP recognizes a pair of FLP recombinase target (FRT) sequences that flank a genomic region of interest.
Which is the temperature optimum for Flp recombination?
Recombination between these sites can lead to excision, inversion, and translocation of DNA in a similar fashion as between loxP sites (For details see Cre-lox blog post ). The first version of FLP discovered has a temperature optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C).
How is the FLP-in system used in research?
The Flp-In System involves introduction of a Flp Recombination Target (FRT) site into the genome of the mammalian cell line of choice. An expression vector containing your gene of interest is then integrated into the genome via Flp recombinase-mediated DNA recombination at the FRT site. For Research Use Only.
What is the difference between C re-LOX and Flp-FRT?
The FLP-FRT system The FLP-FRT system is similar to the C re-lox system and is becoming more frequently used in mouse-based research. It involves using flippase (FLP) recombinase, derived from the yeast Saccharomyces cerevisiae (Sadowski 1995). FLP recognizes a pair of FLP recombinase target (FRT) sequences that flank a genomic region of interest.