Site-specific recombination is also an important process that viruses, such as bacteriophages, adopt to integrate their genetic material into the infected host. Two factors have been shown to affect the efficiency of Cre's excision on the lox pair. Cre, a 38-kDa recombinase from bacteriophage P1 that resolves genome dimers into monomers ( 1, 2 ), is one of the simplest and best understood of known recombinases. The schematic below shows the three types of rearrangements: inversion, deletion and translocation. How do I prepare and deposit my plasmids? Please acknowledge the Expressed in fibroblasts, TFP and Cre-ERT2 - Tamoxifen inducible. Can Cre Recombinase be used to linearize a BAC containing a loxP site? Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. patience as we work to enhance our plasmid and sequence displays. Independently, Joe Z. Tsien has pioneered the use of Cre-loxP system for cell type- and region-specific gene manipulation in the adult brain where hundreds of distinct neuron types may exist and nearly all neurons in the adult brain are known to be post-mitotic. In its lytic state, once its viral genome is injected into the host cell, viral proteins are produced, virions are assembled, and the host cell is lysed to release the phages, continuing the cycle. MeSH Systems, Research Migration of the branch point to the second cleavage site would then somehow trigger the second half of the pathway. The sequence clipboard box will appear with the top strand sequence of the selected region. pCMV6-Entry-Cre Sequence and Map - SnapGene File contains the nucleotide sequence and annotated features in GenBank flat file format. Site-specific recombination (SSR) involves specific sites for the catalyzing action of special enzymes called recombinases. The site is secure. Cre Recombinase - an overview | ScienceDirect Topics Lox sites are directional and the two sites joined by the tetramer are parallel in orientation. The enzyme's unique and specific recombination system is exploited to manipulate genes and chromosomes in a huge range of research, such as gene knock out or knock in studies. Initiation of site-specific recombination begins with the binding of recombination proteins to their respective DNA targets. The components of this recombination system were elucidated using deletion mutagenesis studies. If the 5 hydroxyl groups attack the 3-phosphotyrosine linkage one pair of the DNA strands will exchange to form a Holliday junction intermediate. the plasmid map, sequence, and perform additional sequence analysis. High-resolution specificity profiling and off-target - Nature Metzger, D. and Feil, R (1999). J Vis Exp. Cre-ERT2 - Tamoxifen inducible; Gateway entry vector, Cre expressed at low levels to reduce toxicity; See PI page for other recombinases that are less toxic in Drosophila, Cre-EGFP fusion; Tet inducible - rrTA expression driven by mouse Nkx cardiac enhancer and promoter fragment, N-terminal component of the Co-InCre system, C-terminal component of the Co-InCre system, Cre-ERT2; Targeting vector for Nanog locus, Cre, KASH-tagged EGFP, and sgRNA expression, DHFR-destabilized Cre; targeting vector for Rasgrf2, Cre-ERT2 - Tamoxifen inducible; Targeting vector, Cre fused to supernegatively charged GFP variant, Cre fused to the human Estrogen Binding Domain (EBD), Cre with a 25 nucleotide extracellular vesicle targeting sequence and CFP, mCherry and Cre expression in newborn neurons, GFP and Cre expression in newborn neurons, Cre recombinase dependent on GFP (CRE-DOG), iCre with MCS for inserting promoter, WPRE, CreER expression and tetracyclin-dependent transgene/shRNA expression, mCherry and Cre; expressed in excitatory neurons, For in vitro transcription of Cre or to recombine into BAC, TFP and Cre-ERT2 - Tamoxifen inducible. Depending on the construct, Cre may activate or repress gene expression. During site-specific DNA recombination, which brings about genetic rearrangement in processes such as viral integration and excision and chromosomal segregation, these recombinase enzymes recognize specific DNA sequences and catalyse the reciprocal exchange of DNA strands between these sites. Have questions about your order, deposit, or a plasmid? (A) An overview of Cre-loxP system. The Cre-lox system is used as a genetic tool to control site specific recombination events in genomic DNA. Individual Sequences & Maps. The C domain is similar in structure to the domain in the Integrase family of enzymes isolated from lambda phage. The P1 plasmid is relatively large (90Kbp) and hence exists in a low copy number - usually one per cell. sequence location of the cut. OMP targeting vector with Cre FNF (neo-selectable marker flanked by FRT sites), M71 targeting vector with IRES CreFNF (neo-selectable marker flanked by FRT sites), Cre, Puro resistance and miRNA expression. What do I need to know about the customs and importation process for my country? The schematic below shows the three types of rearrangements: inversion, deletion and translocation. Sequence. It is implemented both in eukaryotic and prokaryotic systems. The sequence clipboard box will appear with the top strand sequence of the selected region. These sites, known as loxP sequences, contain specific binding sites for Cre that surround a directional core sequence where recombination can occur. Commonly used gene replacement strategies introduce selectable markers into the genome to facilitate selection of genetic mutations that may cause growth retardation. Fax: 978-921-1350 "Site-specific recombination of DNA in eukaryotic cells", "Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae", "Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1", "Tissue- and site-specific DNA recombination in transgenic mice", "T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination", "Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting", "Subregion- and cell type-restricted gene knockout in mouse brain", "The essential role of hippocampal CA1 NMDA receptor-dependent synaptic plasticity in spatial memory", http://www.neuroscienceblueprint.nih.gov/factSheet/CreDriver.htm, "Skeletal and CNS defects in Presenilin-1-deficient mice", "Deficient neurogenesis in forebrain-specific presenilin-1 knockout mice is associated with reduced clearance of hippocampal memory traces", "Yeast two-hybrid interaction partner screening through in vivo Cre-mediated Binary Interaction Tag generation", "Cyclization recombinase [Escherichia coli] - Protein - NCBI", "Targeted integration of DNA using mutant lox sites in embryonic stem cells", "A high-throughput screen identifying sequence and promiscuity characteristics of the loxP spacer region in Cre-mediated recombination", "A genetic strategy for stochastic gene activation with regulated sparseness (STARS)", "Engineering mouse chromosomes with Cre-loxP: range, efficiency, and somatic applications", "Non-parallel recombination limits Cre-LoxP-based reporters as precise indicators of conditional genetic manipulation", "Genetically engineered mouse models in cancer research", "Spontaneous recombinase activity of Cre-ERT2 in vivo", "Epigenetic Regulation of Vascular Smooth Muscle Cells by Histone H3 Lysine 9 Dimethylation Attenuates Target Gene-Induction by Inflammatory Signaling", "Extensive Proliferation of a Subset of Differentiated, yet Plastic, Medial Vascular Smooth Muscle Cells Contributes to Neointimal Formation in Mouse Injury and Atherosclerosis Models", "Previously differentiated medial vascular smooth muscle cells contribute to neointima formation following vascular injury", "KLF4-dependent phenotypic modulation of smooth muscle cells has a key role in atherosclerotic plaque pathogenesis", "Lineage tracing of cells involved in atherosclerosis", "Disease-relevant transcriptional signatures identified in individual smooth muscle cells from healthy mouse vessels", Introduction to Cre-lox technology by the "Jackson Laboratory", https://en.wikipedia.org/w/index.php?title=Cre-Lox_recombination&oldid=1156566276, This page was last edited on 23 May 2023, at 14:02. Purification and properties of the Cre recombinase protein", "Crystal structure of a wild-type Cre recombinase-loxP synapse reveals a novel spacer conformation suggesting an alternative mechanism for DNA cleavage activation", "The Cre recombinase cleaves the lox site in trans", "Genetically engineered mouse models in cancer research", "Spontaneous recombinase activity of Cre-ERT2 in vivo", "Mutants of Cre recombinase with improved accuracy", https://en.wikipedia.org/w/index.php?title=Cre_recombinase&oldid=1135615154, Short description is different from Wikidata, Articles with German-language sources (de), Creative Commons Attribution-ShareAlike License 4.0, This page was last edited on 25 January 2023, at 19:59. Store, search, and share your sequences, files and maps; Download SnapGene Viewer. Cre recombinase: The universal reagent for genome tailoring Editing, Cloning Leading primers are indicated on the first line of each sequence. Kozak consensus sequence was added before the start ATG.Myc-tag was added at the C-terminus of Cre. These studies showed that a P1 gene product and a recombination site were both required for efficient recombination events to occur. Addgene: CRE recombinase email or call1-800-NEB-LABS. The Cre recombinase of the P1 bacteriophage belongs to the integrase family of site-specic recombinases. The P1 phage is a temperate phage that causes either a lysogenic or lytic cycle when it infects a bacterium. PMC The protein will stay in this location in its inactivated state until tamoxifen is given. Does Addgene accept orders by fax, phone or email? In the lysogenic cycle the phage genome replicates with the rest of the bacterial genome and is transmitted to daughter cells at each subsequent cell division. If you run into any problems registering, depositing, or ordering please contact us at [emailprotected] Mouse Cre-LoxP system: general principles to determine tissue-specific Expressed in chondrocytes, mCherry and Cre-ERT2 - Tamoxifen inducible; Targeting vector for Pax7, Venus and Cre-ERT2 with MCS for inserting promoter, Emerald and Cre-ERT2 with MCS for inserting promoter, TFP and Cre-ERT2 with MCS for inserting promoter, tdTomato and Cre-ERT2 with MCS for inserting promoter, Venus, Cre-ERT2, targeting vector with MCS for homology arms, Emerald, Cre-ERT2, targeting vector with MCS for homology arms, TFP, Cre-ERT2, targeting vector with MCS for homology arms, tdTomato, Cre-ERT2, targeting vector with MCS for homology arms, GAG-Cre fusion; for the production of VLPs loaded with CRE protein, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, unskippable linker) - Tamoxifen inducible, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, GSAx9 linker) - Tamoxifen inducible, iCre-ERT2 (PAPGSTMA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PA N-terminus) - Tamoxifen inducible, CreLite - Red light-inducible Cre; N terminus Cre fused to PIF6, CreLite - Red light-inducible Cre; C terminus Cre fused to PhyBdelta, CreLite system components, PhyBdeltaCreC and PIF6CreN, in middle entry vector (Tol2 kit), CreLite; Tol2 destination vector with mTagBFP2, CreLite system components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer vector, CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector, Cre recombinase split with Vivid photodimers, Cre-ERT2 - Tamoxifen inducible; AAV donor vector, iCre-ERT2 (PAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSAS N-terminus) - Tamoxifen inducible, N-terminal Cre component split with Vivid photodimers, C-terminal Cre component split with Vivid photodimers, Split Cre fused to nuclear localized wild-type VVD, Retinal ganglion cell-specific expression of Cre, Cre activates shRNA expression, removal of EGFP; See also similar plasmids, Cre turns off shRNA expression, removal of EGFP; See also similar plasmids, Cre activates gene of interest by removing Stop sequences; Puro selection.
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cre recombinase sequence