What is FRT sequence?
FRT sites are used often (at least in Drosophila) for inducing deletions or “flipping out” of markers in transgenic constructs. When there are two FRTs sequences in tandem, after inducing flippase the DNA sequence that is between these two sites will be deleted.
What is FLP-FRT used for?
In genetics, Flp-FRT recombination is a site-directed recombination technology, increasingly used to manipulate an organism’s DNA under controlled conditions in vivo.
What organism does the FRT FLP system originated from?
The FLP-FRT system. The FLP-FRT system is similar to the Cre-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).
What is pcDNA5?
pcDNA5/TO is a 5.7 kb expression vector designed for use with the T-REx System (Cat. The pcDNA5/TO vector allows tetracycline-regulated expression of the gene of interest in mammalian host cells expressing the Tet repressor (TetR) from the pcDNA6/TR vector (Catalog no. V1025-20).
What is FRT Drosophila?
Fly Board Election The FRT entry in FlyBase represents the wild-type target site for the FLP recombinase encoded by the Saccharomyces cerevisiae 2μ plasmid. It is expected to be compatible with any engineered derivative of FLP in which the target site specificity of the recombinase has not been altered.
What are Cre-lox mice?
The Cre/lox system is one of the most powerful and versatile tools developed for mouse genetics. It gives mouse researchers sophisticated control over the location and timing of gene expression. Cre/lox is usually used to make knockout alleles, but it can also be used to activate gene expression.
Is Cre reversible?
All recombination events mediated by FLP or Cre are reversible. Because recombination can only occur between target sites of the same sequence in FLEx vectors, the DNA sequence that should be inverted is flanked by two pairs of target sites that differ in their sequence.
What is a Cre reporter mice?
Abstract. The Cre/loxP system is a strategy for controlling temporal and/or spatial gene expression through genome alteration in mice. As successful Cre/loxP genome alteration depends on Cre-driver mice, Cre-reporter mice are essential for validation of Cre gene expression in vivo.
How does Cre-LOX work?
Overview. Cre-Lox recombination involves the targeting of a specific sequence of DNA and splicing it with the help of an enzyme called Cre recombinase. Cre-Lox recombination is commonly used to circumvent embryonic lethality caused by systemic inactivation of many genes.
How does Cre LOX system work?
Concerning the mechanism of Cre-loxP system, a single Cre recombinase recognizes two directly repeated loxP site, then the Cre excises the loxP flanked (floxed) DNA, thus creating two types of DNA with circular, excised and inactivated gene Y (Figure 1A).
How does the pcdna™5/frt plasmid work?
The pcDNA™5/FRT vector contains a single FRT site immediately upstream of the hygromycin resistance gene for Flp recombinase-mediated integration and selection of the pcDNA™5/FRT plasmid following cotransfection of the vector (with pOG44) into Flp-In™ mammalian host cells.
How is the pog44 plasmid cotransfected in FLP-in™?
The pOG44 plasmid and the pcDNA5/FRT vector containing your gene of interest are cotransfected into the Flp-In™ host cell line.
What is Flp recombinase target (FRT) sequence?
FLP recognizes a pair of FLP recombinase target (FRT) sequences that flank a genomic region of interest. Araki K, Imaizumi T, Okuyama K, Oike Y, Yamamura K. 1997. Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. J Biochem (Tokyo) 122:977-82.
What is the FLP-FRT system?
The FLP-FRT system. The FLP-FRT system is similar to the Cre-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.