What is the effect of FLP on FRT?
When the plants were exposed to heat-shock, FLP-induction led to the excision of the FRT-flanked vector, effectively moving the GUS gene directly downstream of the CaMV 35S promoter.
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 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 FLPo?
FLPo is a codon-optimized FLP version first described by Raymond and Soriano (2007). Its amino acid sequence is identical to that of FLPe but the codon usage was altered to improve expression in mammalian cells. It appears to be at least 10 fold more efficient than FLPe.
What is Cre mouse line?
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.
What is conditional deletion?
Conditional gene knockout is a technique used to eliminate a specific gene in a certain tissue, such as the liver. It differs from traditional gene knockout because it targets specific genes at specific times rather than being deleted from beginning of life.
What is FRT Drosophila?
The FLP/FRT system is a site-directed recombination technology based on the targeting of a recombination enzyme (flipase – FLP) to specific DNA regions designated as flipase recognition target (FRT) sites.
Is Cre recombination 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 floxed allele?
In genetics, floxing refers to the sandwiching of a DNA sequence (which is then said to be floxed) between two lox P sites. The terms are constructed upon the phrase “flanking/flanked by LoxP”. Floxing a gene allows it to be deleted (knocked out), translocated or inverted in a process called Cre-Lox recombination.
Can FLP-mediated recombination occur between pcdna5/frt and the integrated FRT site?
Flp-mediated recombination between the FRT site on pcDNA5/FRT and the integrated FRT site in the Flp-In™ host cell line will only occur if the pcDNA5/FRT plasmid is circularized. The pOG44 plasmid should be circularized to minimize the possibility of the plasmid integrating into the genome.
What is FLP-FRT recombination?
From Wikipedia, the free encyclopedia 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 is the role of the FLP recombinase during cotransfection?
Upon cotransfection, the Flp recombinase expressed from pOG44 mediates a homologous recombination event between the FRT sites (integrated into the genome and on pcDNA5/FRT) such that the pcDNA5/FRT construct is inserted into the genome at the integrated FRT site (see the figure below).
What is short flippase recognition target (FRT) recombination?
It is analogous to Cre- lox recombination but involves the recombination of sequences between short flippase recognition target ( FRT) sites by the recombinase flippase ( Flp) derived from the 2 µ plasmid of baker’s yeast Saccharomyces cerevisiae . The 34bp minimal FRT site sequence has the sequence