...the core sequence gives the loxP site directionality, and the canonical loxP sequence is ATAACTTCGTATA-... a single loxP site are recognized and bound by a Cre protein, forming a dimer. The two loxP sites then... location and orientation of the loxP sites: Inversion: If the loxP sites are on the same DNA strand ... putting the loxP sites on either side of a gene (called “floxing”, for “flanked by loxP”), will permit...potential limitation of using loxP sites is the inability to tightly control which loxP sites recombine if more... the P1 bacteriophage: the Cre recombinase and a loxP recognition site. The P1 bacteriophage uses these... intra- and inter-molecular recombination at the loxP recognition sites. A key advantage of the system...

...lox511 sites, not with loxP sites. Alternatively, you could use a pair of loxP sites and a pair of FRT...between the identical loxP or lox511 sites (Figure 1, bottom). Since only one loxP and lox511 site will...with one loxP site and one lox511 site in the same orientation, and placing the additional loxP and lox511... our Plasmids 101 series, we learned how the Cre-loxP recombination system can be used to induce site-...events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert, translocate...fragment. The availability of both wild-type and mutant loxP sites has allowed scientists to leverage this system...specific recombination events: Cre recombinase binds loxP sites, while FLP binds FRT sites. When a DNA sequence...

...resistance gene through the insertion of flanking LoxP sequences, you can remove the gene through the addition...design your targeting vector such that a set of three LoxP sites flank the resistance gene and the targeted... been replaced with the same sequence flanked by LoxP sites while the resistance cassette has been placed... marker using Cre recombinase. Since there are 3 loxP sites, recombination can occur in multiple ways....floxed, as seen in line 4 of Figure 4. Since the loxP sites are located in intronic regions, this gene...using a repair template. These templates can include loxP sites to create conditional floxed alleles. Although...

...recombination between those sites. FRT sites and loxP sites differ at the nucleotide level but share an...translocation of DNA in a similar fashion as between loxP sites (For details see Cre-lox blog post). Optimizing...FLP-FRT systems in your experiments. Mutated FRT and loxP sites in Flex Vectors – inversion only once All ...Whereas the excision of a piece of DNA flanked by loxP/FRT sites is favored over its reintroduction, inversion... sequence and should therefore be prevented. The loxP and FRT target sites have been engineered to avoid...Cre-ERT2 recombinase, leading to recombination of the loxP sites and thereby to the gene knockout. In this ...

...sandwiched between 2 loxp sites and is excised by Cre (Figure 1). This results in one loxp site and one lox2272...unused (loxp) sites will be flanked by two lox sites now in the same orientation. Next, the loxp site is..., but they are comprised of different sequences (loxp and lox2272) and can not recombine with each other...system it binds one pair of lox sites, let's say the loxp pair, and flips the transgene into the correct orientation...lox2272 site on either end of the transgene. Since loxp and lox2272 can not recombine, this ensures that...

...target 2 loxP sites to intronic regions 0.5 - 0.8 kb apart. In the example below, we’ll add loxP sites for... and two repair templates in order to insert two loxP sites. Since two double-strand breaks are made, ...variety of other undesired outcomes, including one loxP site insertion and one NHEJ-repaired indel, often...homology on each side. For optimal repair, place the loxP sites precisely at the cut site for each guide. ...

...these deletions are defined using 2 loxP variants (only identical loxP sites can mediate recombination.)...recombination events, and the synthesis of alternative loxP sites to specify distinct patterns of recombination...exclusive, as either deletion removes one of the other loxP site variants, preventing further recombination....proteins, with recombination events specified by three loxP variants, but the principles remain the same. The...

...inducible yet permanent gene excision. Essentially, LoxP sites are knocked in flanking a gene of interest...time-tested system but does require knock-in of the LoxP sites, which is more difficult than a simple knock...out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little while Transient...

...recombination of DNA sequences located between two loxP sites. The loxP sites are present at both ends of a gene...recombination. To generate a conditional knockout mouse, loxP sites are inserted directly into the genome to flank...

...genetic recombination in mice expressing both Cre and LoxP occurs. But how are these mice made to begin with...want to genotype your mice for expression of Cre, loxP, and your gene of interest. To ensure that the effects...control should be littermates that express Cre but not loxP. With this two-part guide to mouse modeling now ...

...Cre may induce recombination at pseudo- or cryptic loxP sites (estimated to occur at a frequency of 1.2 ...458-65. doi:10.1007/s004270100167. PubMed. Cryptic loxP sites in mammalian genomes: genome-wide distribution...

...recombinase recognizes 34 bp loxP sites, and the orientation and location of the loxP sites determines how the... putting the loxP sites on either side of a gene (called “floxing”, for “flanked by loxP”), will permit...IRES-EGFP Mammalian Costantini 32702 pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE Cre activates your gene fused...more colors. Mammalian Zeng 24334 hsp70l-loxP-mCherry-STOP-loxP-H2B-GFP_cryaa-cerulean Heat-inducible reporter... reporter Lentiviral Geijsen 65726 pLV-CMV-LoxP-DsRed-LoxP-eGFP Switches from DsRed to eGFP expression...expression Mammalian Green 32145 pJFRC172-10XUAS-loxP>-dSTOP-loxP>-myr::GFP Cre dependent myr-GFP expression...recombinases that recognize specific mutant loxP sites, VloxP and SloxP sites respectively. Split Cre: Cre recombinase...

...easy to remove after injection, they added flanking loxP sites and a heat-shock inducible Cre to generate...schematic below.) After self-excision, the resulting loxP scar is located in this synthetic intron, and only...

...technology (Cre/loxP) to remove the donor cassette. While this method can be effective, a loxP site is left...Grégoire and M. Kmita, “Recombination between inverted loxP sites is cytotoxic for proliferating cells and provides...

...recombination at defined recognition sites (i.e., loxP and FRT, respectively) (Abremski and Hoess, 1984...two sets of incompatible recognition sites (e.g., loxP and lox511) in opposite orientations. Since Cre ... Van Duyne, Gregory D. "A structural view of cre-loxp site-specific recombination." Annual review of biophysics...

...puromycin or screen for GFP Bartel 32702 pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE MSCV Conditional overexpression...

...Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV vectors are common approaches...

...stochastic multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein...

...series of primary cell types using Cre and various loxP reporters. In both mouse and human embryonic stem...

...transgenes that are flanked by loxP sites. While there is only a small loxP scar behind at the site of excision...