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Cre Recombinase protein delivered by cell-derived nanovesicles Cre gesicles tech note
Home › Products › Gene function › Gene editing › Cre recombinase › Cre Recombinase Gesicles

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Cre Recombinase protein delivered by cell-derived nanovesicles Cre gesicles tech note

Cre Recombinase Gesicles

Deliver Cre recombinase protein to mammalian cells in the absence of nucleic acids (i.e., no plasmid or virus is required), rapidly and efficiently. These gesicles are cell-derived nanovesicles containing concentrated Cre recombinase protein that, when applied to your target cells, fuse with the plasma membrane and discharge into the lumen of the cell.

Deliver Cre recombinase protein to mammalian cells in the absence of nucleic acids (i.e., no plasmid or virus is required), rapidly and efficiently. These gesicles are cell-derived nanovesicles containing concentrated Cre recombinase protein that, when applied to your target cells, fuse with the plasma membrane and discharge into the lumen of the cell. The enzyme is transported to the nucleus via its nuclear localization signal where it can carry out Cre-mediated recombination between loxP sites. Since there is no coding gene present, there is no risk of sustained Cre recombinase expression in your target cells, thus limiting the chance of unintended recombination events.

What is Cre recombinase?

Cre recombinase is a site-specific recombinase isolated from P1 bacteriophage that catalyzes the recombination between two 34­-bp loxP sites. The enzyme is widely used for knock-out and knock-in studies: any DNA sequence located between two tandem repeats of loxP can be excised to create a deletion mutant, or a plasmid containing a single loxP site can undergo site-specific integration into a genome containing loxP to create an insertion mutant.

What are the benefits of gesicle technology?

  • Far simpler to use than either plasmid or viral gene delivery. Simply apply 10–20 µl of the gesicles to your target cells; no preparation or pre-treatment is required.
  • Flox your cells on demand: Cre recombinase is present when you need it and gone when you don’t.
  • Can be used with many cell types, including dividing and non-dividing cells, primary cells, and cell lines.
  • Ensure efficient delivery and high activity of Cre recombinase (expand the Details tab in the product table to see data images). When using gesicle delivery, there is no delay for transcription/translation: the protein is present and active as soon as you apply the gesicles to your cells.
  • Since there is no coding gene present, there is no risk of sustained Cre recombinase expression in your target cells. It is present when you need it and gone when you don’t, so you avoid unwanted recombination events.
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Cat. # Product Size Price License Quantity Details
631449 Cre Recombinase Gesicles 200 uL USD $586.00

License Statement

ID Number  
83 The Living Colors mCherry Monoclonal Antibody has been obtained from National University of Shimane.
225 Gesicle Technology. This product and its use are the subject of European patents and patent applications related to EP09306091, owned by Inserm Transfert. For research use only – not for therapeutic or diagnostic use in humans. No rights granted other that the right to use for research purposes.

In addition, Inserm Transfert is the owner of European patent application EP09306092.9 relating to the use of microvesicles for cell engineering purposes. The products should not be used to develop, make, have made, use and sell or otherwise distribute any product, composition, method, service or process for commercial services that would infringe said patent application, unless a license is negotiated with Inserm Transfert, at 7 rue Watt, 75013 Paris, France (www.inserm-transfert.fr).
405
This Product is protected by one or more patents from the family consisting of: US9593356, CH3008192, People's Republic of China Patent ZL201480032802.9, DE602014050225.3, US10793828, US11643635, EP3008192, FR3008192, UK3008192, JP6525971, JP6771068, CA2905229 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

259 This Product is protected by one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

Cre Recombinase Gesicles are cell-derived nanovesicles used to deliver active Cre recombinase protein directly into target cells. When they are applied to your target cells, the gesicles fuse with the plasma membrane and deliver a temporary dose of Cre recombinase, which facilitates recombination at loxP sites contained within the cell. As an added benefit, Cre Recombinase Gesicles contain CherryPicker protein, a membrane-targeted version of the mCherry fluorescent protein that helps to confirm the delivery of the Cre recombinase to your target cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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293T cells are used to produce Cre Recombinase Gesicles, which allow rapid and efficient genome modification in the target cells of your choice

293T cells are used to produce Cre Recombinase Gesicles, which allow rapid and efficient genome modification in the target cells of your choice
293T cells are used to produce Cre Recombinase Gesicles, which allow rapid and efficient genome modification in the target cells of your choice. Nanovesicle-inducing glycoproteins stimulate gesicle formation from the 293T cell line. iDimerize technology incorporates Cre recombinase into the gesicles and associates it with CherryPicker red fluorescent protein. After the gesicles pinch off from the producer cells, they fuse with the plasma membrane of the target cells and release Cre recombinase into the cell. The CherryPicker protein provides convenient visualization of delivery to your cells.

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ZsGreen1 reporter assay for efficiency of Cre Recombinase Gesicles with various cell lines

ZsGreen1 reporter assay for efficiency of Cre Recombinase Gesicles with various cell lines
ZsGreen1 reporter assay for efficiency of Cre Recombinase Gesicles with various cell lines. pLVX-LoxP-ZsGreen1 contains ZsGreen1 cDNA separated from the EF1a promoter by a floxed stop cassette. In the presence of Cre recombinase, the stop cassette is removed, permitting high-level expression of ZsGreen1. All cell lines were transduced with this vector, followed by selection in puromycin to create stable lines. These lines were then exposed to a dilution series of recombinase-containing gesicles, centrifuged at 1,000 x g for 30 min at 32°C, and incubated for an additional 3 hr at 37°C. After 3 hr, media was exchanged and cells were permitted to grow for an additional 48 hr, at which time they were analyzed by FACS for ZsGreen1 expression.

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LacZ Reporter Assay for Cre Recombinase Gesicles

LacZ Reporter Assay for Cre Recombinase Gesicles
LacZ Reporter Assay for Cre Recombinase Gesicles. Panel A. Gesicle effectiveness was tested using a reporter cell line in which the lacZ gene is only expressed following removal of a stop codon by Cre recombinase activity. Cells were either transfected for 6 hr with a plasmid expressing Cre recombinase, or treated with 20 µl Cre Recombinase Gesicles for 3 hr. 24 hr after treatment, cells were stained for lacZ expression using the Beta Galactosidase Staining Kit (Cat. No. 31780). Panel B. The high concentration of blue cells in the center image indicate high recombinase activity. Red fluorescence in the right-hand image provides easy visualization of gesicle delivery to target cells.

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Visualization of ZsGreen1 reporter assay for Cre Recombinase Gesicles in various cell lines

Visualization of ZsGreen1 reporter assay for Cre Recombinase Gesicles in various cell lines
Visualization of ZsGreen1 reporter assay for Cre Recombinase Gesicles in various cell lines. pLVX-LoxP-ZsGreen1 contains ZsGreen1 cDNA separated from the EF1a promoter by a floxed stop cassette. In the presence of Cre recombinase, the stop cassette is removed, permitting high-level expression of ZsGreen1. All cell lines were transduced with this vector, followed by selection in puromycin to create stable lines. These lines were then exposed to a dilution series of recombinase-containing gesicles, centrifuged at 1,000 x g for 30 min at 32°C, and incubated for an additional 3 hr at 37°C. After 3 hr, media was exchanged and cells were permitted to grow for an additional 48 hr, at which time they were imaged by fluorescence microscopy for ZsGreen1 expression.

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631449: Cre Recombinase Gesicles

631449: Cre Recombinase Gesicles

More Information

Applications

  • For knockout and knockin studies
  • Limit and control site-specific recombination events including deletions, insertions, translocations, and inversions

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.






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Tools for delivering Cre recombinase

Rapid, efficient Cre recombinase delivery for genome modification

See how gesicle technology outperforms plasmid transfection for efficient delivery of Cre recombinase in most cell types.

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