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Cat. No.: TCRE-200 (for 200μg)

Cat. No.: TCRE-1k (for 1mg)

Cat. No.: TCRE-5k (for 5mg)

Description

TAT-Cre Recombinase is a Cre Recombinase recombinant protein that contains an N-terminal His-tag, a cell-penetrating peptide TAT (Transactivator of Transcription), and a Nuclear Localization Signal (NLS). This product is commonly used to incubate with cultured cells, enabling site-specific recombination of DNA at loxP sites within the cells.

Cre Recombinase is a type I topoisomerase derived from the Escherichia coli bacteriophage P1. It is also a tyrosine recombinase that recognizes 34 bp loxP sites, which consist of two 13 bp inverted repeats (ATAACTTCGTATA and TATACGAAGTTAT) flanking an 8 bp spacer region (NNNTANNN). Cre Recombinase catalyzes the recombination of DNA between loxP sites. The recombination products differ depending on the position and relative orientation of the loxP sites. DNA containing single loxP sites will fuse; DNA between two loxP sites in the same direction will be excised as a circular fragment, while DNA between two oppositely oriented loxP sites will be inverted.

The cell-penetrating peptide TAT is derived from a positively charged peptide segment of approximately 10 amino acids from the transactivator of transcription factor, which is essential for the replication and gene expression of HIV-1. Research has shown that TAT fusion proteins can be internalized by cells via caveolar endocytosis, independent of clathrin-mediated endocytosis. This allows various molecules fused with TAT (including peptides, proteins, antisense oligonucleotide strands, magnetic nanoparticles, and liposomes) to penetrate cell membranes, making them suitable for many applications.

Cre Recombinase has widespread application value in genetic modification of DNA both in vitro and in vivo, but one of its limitations is the difficulty in expressing and delivering the enzyme with stable activity in mammalian cells. Common approaches to address this issue include transient transfection of DNA vectors expressing Cre Recombinase, although this often results in low transfection efficiency and unstable or low recombination efficiency; and viral transduction, which is more effective but carries the risk of insertional mutagenesis. We have developed a recombinant fusion protein of cell-permeable TAT-Cre Recombinase that can be directly delivered into mammalian cells. Assisted by the NLS, the fusion protein rapidly enters the nucleus and catalyzes efficient recombination, with precise control over dosage and duration, avoiding the integration of exogenous DNA into the genome, which could affect experimental results.

Source

TAT-Cre Recombinase is obtained through the recombination, expression, and purification of Escherichia coli (E. coli).

Use

Catalyzes site-specific recombination of sequences at loxP sites in vivo.

Concentration

4 mg/ml (approximately 100 μM).

Purity

Greater than 95%, as determined by SDS-PAGE electrophoresis.

Enzyme Storage Solution

50 mM NaH2PO4, 500 mM NaCl, 5 mM Tris, pH 8.0, 10% (v/v) Glycerol.

Precautions

• High concentrations of TAT-Cre Recombinase can be toxic to cells. It is recommended to conduct preliminary experiments to determine the optimal working concentration. Note that cell toxicity is a normal phenomenon, and a small portion of cells surviving often indicates better recombination results.

• This product is intended for scientific research use only by trained professionals and must not be used for clinical diagnosis or treatment, food or drugs, or stored in a regular household.

• For your safety and health, please wear a lab coat and disposable gloves during handling.

Storage

Store at -80ºC, valid for one year.

Related: Cre Recombinase