| Cat # |
EG-1001 (old catalog #: RP-7) |
| Discount |
$100 Discount Code for Your First Order of 1 mg Cre Recombinase: CreDiscount
Discount for switching from competitors' product: $150. Email to Tech Support crm@excellgen.com for discount code. |
| Description |
Cre recombinase, often abbreviated to Cre, is a Type I topoisomerase from P1 bacteriophage that catalyzes site-specific recombination of DNA between loxP sites. This enzyme does not require any energy cofactors, and Cre-mediated recombination quickly reaches equilibrium between substrate and reaction products. The loxP recognition element is a 34 base pair (bp) sequence composed of two 13 bp inverted repeats flanking an 8 bp spacer region which confers directionality. Recombination products are dependent on the location and relative orientation of the loxP sites. Two DNA species containing single loxP sites will be fused whilst DNA between loxP sites in the same orientation will be excised in circular form and DNA between opposing loxP sites will be inverted with respect to the rest of the DNA.
Cre recombinase is used as a tool to modify genes and chromosomes. In this approach the Cre recombinase is used to delete a segment of DNA flanked by LoxP sites (aka 'floxed') in an experimental animal. It has been used to generate animals with mutations limited to certain cell types (tissue-specific knockout) or animals with mutations that can be activated by drug administration (inducible knockout) in a number of transgenic species. The availability of transgenic lines with tissue specific or inducible Cre expression permits researchers to inactivate or activate a gene of interest simply by breeding a floxed animal to pre-existing Cre-transgenics. One example of an inducible Cre recombinase system is the Cre-ER (ER = Estrogen Receptor) system in which intraperitoneal injection of tamoxifen will cause dose-dependent excision of the floxed site (i.e. will inactivate the gene of choice).
Recombinant Cre recombinase (TAT-Cre) was purified from an E. coli strain carrying an engineered plasmid encoding enhanced form of Cre Recombinase from bacteriophage P1. This Cre recombinase has an N-terminal 6XHis tag, a Tat peptide (GRKKRRQRRRPPAGTSVSL) and an NLS sequence (PKKKRKV). HTNC is the most effective protein in transduction (in vivo) and subsequent recombination compared to other forms of Cre recombinases, e.g., HNC, TCH6, HC, HNCM, CH. Incubation of fibroblast reporter cells with 1 μM HTNC for 1 to 2 hours can result in tranduction of 60 ~ 90% of the cells*. Addition of 100 μM chloroquine to culture medium may further enhance transduction and recombination.
* Use this page to calculate concentration required for cultured cells: http://www.excellgen.com/pub/mw_2_moles.html |
| Applications |
- In vitro LoxP recombination for subcloning or vector/clone engineering
- Transduction into cultured cells including stem cells ex Vivo
|
| Properties |
Concentration: 2, 5, or 10 mg/ml. Molecular Weight: 43 kDa
QC: Tat-Cre induced 80~100% recombination efficiencies in HEK293T- Cre reporter cells. HEK293T-cells were transfected with reporter plasmid DNA: LoxP-RFP-Stop-LoxP-GFP, then add 1 μM purified Tat-Cre protein.
Fig 1: 0: before adding Tat-Cre; 1 to 5: 1 to 5 days after transduction with Tat-Cre.
Endotoxin Levels: < 0.1 EU/μg Purity: >98% by SDS-PAGE and HPLC analysis. |
| Unit Defintion |
A standard of 100 Units is defined as the amount of TAT-CRE (μg) in 1.0 mL of tissue culture medium that is required to induce 50% GFP expression in a HEK 293T reporter cell line assay. |
| Stability |
2 years at -80 oC. 6 months at -20 oC. Upto 3 weeks at 4 oC |
| Citations |
1, Critical Roles of E2F3 in Growth and Musculo-skeletal Phenotype in Mice. Int J Med Sci 16 12 1557-1563 10.7150/ijms.39068
2, Mutationally-activated PI3'-kinase-α promotes de-differentiation of lung tumors initiated by the BRAFV600E oncoprotein kinase. Elife 8 10.7554/eLife.43668
3, The Notch signaling pathway promotes basophil responses during helminth-induced type 2 inflammation. J Exp Med 216 6 1268-1279 10.1084/jem.20180131
4, Patterned human microvascular grafts enable rapid vascularization and increase perfusion in infarcted rat hearts. Nat Commun 10 1 584 10.1038/s41467-019-08388-7
5, Upregulation of Ets1 expression by NFATc2 and NFKB1/RELA promotes breast cancer cell invasiveness. Oncogenesis 7 11 91 10.1038/s41389-018-0101-3
6, Non-canonical NF-κB Antagonizes STING Sensor-Mediated DNA Sensing in Radiotherapy. Immunity 49 3 490-503.e4 10.1016/j.immuni.2018.07.008
7, A cleavage product of Polycystin-1 is a mitochondrial matrix protein that affects mitochondria morphology and function when heterologously expressed. Sci Rep 8 1 2743 10.1038/s41598-018-20856-6
8, Generation of a rod-specific NRL reporter line in human pluripotent stem cells. Sci Rep 8 1 2370 10.1038/s41598-018-20813-3
9, PD-1 is a haploinsufficient suppressor of T cell lymphomagenesis. Nature 552 7683 121-125 10.1038/nature24649
10, Human pluripotent stem cell-derived epicardial progenitors can differentiate to endocardial-like endothelial cells. Bioeng Transl Med 2 2 191-201 10.1002/btm2.10062
11, Sequence segregation improves non-covalent protein delivery. J Control Release 254 131-136 10.1016/j.jconrel.2017.03.387
12, Long-term self-renewing human epicardial cells generated from pluripotent stem cells under defined xeno-free conditions. Nat Biomed Eng 1 10.1038/s41551-016-0003
13, Disease Model of GATA4 Mutation Reveals Transcription Factor Cooperativity in Human Cardiogenesis. Cell 167 7 1734-1749.e22 10.1016/j.cell.2016.11.033
14, Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis. Nat Chem Biol 13 1 81-90 10.1038/nchembio.2238
15, Tumor-infiltrating Tim-3+ T cells proliferate avidly except when PD-1 is co-expressed: Evidence for intracellular cross talk. Oncoimmunology 5 10 e1200778 10.1080/2162402X.2016.1200778
16, Saturated Fatty Acids Engage an IRE1α-Dependent Pathway to Activate the NLRP3 Inflammasome in Myeloid Cells. Cell Rep 14 11 2611-2623 10.1016/j.celrep.2016.02.053
17, A new in vitro mouse oligodendrocyte precursor cell migration assay reveals a role for integrin-linked kinase in cell motility. BMC Neurosci 17 7 10.1186/s12868-016-0242-2
18, Fatty Acid Oxidation is Impaired in An Orthologous Mouse Model of Autosomal Dominant Polycystic Kidney Disease. EBioMedicine 5 183-192 10.1016/j.ebiom.2016.01.027
19, Discovery of a non-cationic cell penetrating peptide derived from membrane-interacting human proteins and its potential as a protein delivery carrier. Sci Rep 5 11719 10.1038/srep11719
20, The role of antisense long noncoding RNA in small RNA-triggered gene activation. RNA 20 12 1916-1928 10.1261/rna.043968.113
21, Deficiency of the DNA repair protein nibrin increases the basal but not the radiation induced mutation frequency in vivo. Mutat Res 769 11-16 10.1016/j.mrfmmm.2014.07.001
22, Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase. Transgenic Res 23 1 177-185 10.1007/s11248-013-9764-x
23, T cell-derived IL-17 mediates epithelial changes in the airway and drives pulmonary neutrophilia. J Immunol 191 6 3100-3111 10.4049/jimmunol.1301360
24, Integrin-linked kinase regulates process extension in oligodendrocytes via control of actin cytoskeletal dynamics. J Neurosci 33 23 9781-9793 10.1523/JNEUROSCI.5582-12.2013
25, Defective glucose metabolism in polycystic kidney disease identifies a new therapeutic strategy. Nat Med 19 4 488-493 10.1038/nm.3092
26, Kinase-independent feedback of the TAK1/TAB1 complex on BCL10 turnover and NF-κB activation. Mol Cell Biol 33 6 1149-1163 10.1128/MCB.06407-11
27, Cutting edge: Krűppel-like factor 2 is required for phenotypic maintenance but not development of B1 B cells. J Immunol 189 7 3293-3297 10.4049/jimmunol.1201439 |
| Storage |
-20ºC ~ -80ºC |
| Shipping |
dry ice |