Ordering information| Product | Catalog Number | Amount of reagent | Amount of glucose solution |
| in vivo jetPEI™-Gal | 202-10G | 0.1 ml | 10 ml |
| in vivo jetPEI™-Man | 203-10G | 0.1 ml | 10 ml |
| in vivo jetPEI™-FluoF | 205-10G | 0.1 ml | 10 ml |
| in vivo jetPEI™-FluoR | 206-10G | 0.1 ml | 10 ml |
2009/08/07
in vivo-jetPEI™ derivatives
jetSI™ 10 mM
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Please note that neither glucose solution nor DOPE are included. |
1. Hassani, Z., Lemkine, G.F., Erbacher, P., Palmier, K., Alfama, G., Giovannangeli, C., Behr, J.P. and Demeneix, B.A. (2005) Lipid-mediated siRNA delivery down-regulates exogenous gene expression in the mouse brain at picomolar levels. J Gene Med 7(2): 198-207.
2. Froidevaux M. S., Berg P., Seugnet I., Decherf S., Becker N., Sachs L. M., Bilesimo P., Nygard M., Pongratz I., Demeneix B. A., (2006) The co-chaperone XAP2 is required for activation of hypothalamic thyrotropin-releasing hormone transcription in vivo EMBO Rep (7) 1182.
3. Guissouma H., Froidevaux M. S., Hassani Z., Demeneix B. A., (2006) In vivo siRNA delivery to the mouse hypothalamus confirms distinct roles of TR beta isoforms in regulating TRH transcription Neurosci Lett (406) 240-3.
4. Kumar P., Lee S. K., Shankar P. and Manjunath N. (2006) A single siRNA suppresses fatal encephalllitis induced by two different flaviviruses PLOS Med 3: 0505-1.
5. Cheret, C., Gervais, A., Lelli, A., Colin, C., Amar, L., Ravassard, P., Mallet, J., Cumano, A., Krause, K. H., Mallat, M. (2008).Neurotoxic activation of microglia is promoted by a nox1-dependent NADPH oxidase, J Neurosci 28, 12039.
6. Cakir, I., Perello, M., Lansari, O., Messier, N. J., Vaslet, C. A., Nillni, E. A. (2009) Hypothalamic Sirt1 regulates food intake in a rodent model system, PLoS One 4, e8322.
FectoFly™
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1 ml of FectoFly™ is sufficient to perform 100 to 200 transfections in 6-well plates. For bulk sizes, Please contact us. |
By simple and rapid transient transfection method, FectoFly™ provides high transfection efficiency in insect cells and robust protein production over several days (Fig. 1).
FectoFly™ provides high transfection efficiency in most commonly used insect cells (Fig. 2) allowing efficient protein production.
Figure 2. Comparison of FectoFly™ in various adherent cell lines. Sf9, Sf21, S2 or Tn5 adherent cells were transfected in 24-well plates pCMV-EGFP-Luc plasmid. Luciferase assays were performed 72 h after transfection.
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FectoFly™ allows successful transfection of adherent and suspension insect cell cultures both in the latest generation of syntheticserum-free media (Fig.1, 3 – Sf21 and Sf9) and in the presence of serum (Fig.3 – S2). Transfection efficiency ranges routinely from 30% to 50% and cell viability is above 80%.
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Fig 3. GFP expression in Sf21, Sf9 and S2 cells grown in suspension, 72 h after transfection of pCMV-EGFP-Luc plasmid using FectoFly™. |
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FectoFly™ was compared to other commercially available reagents dedicated to the transfection of insect cells (Fig. 4). As shown for Sf9 cells, the highest protein expression levels were obtained with FectoFly™.
| Transfection of insect cells with FectoFly™ is straightforward. o Fast : 3 steps protocol o Simple: 1 µl of FectoFly™ per µg of DNA, 1:1 ratio o Versatile: efficient in serum-free synthetic media and in the presence of serum
Figure 5. FectoFly™ standard protocol. |
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PEIpro™
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Large sizes are available upon request. Please contact us. |
PEIpro™ has been formulated at 1 mg/mL to fit current standard for transfection with PEI. This concentration eases the transfection protocol optimization. PEIpro protocol is available upon request. Contact the Technical Support.
Positively charged reagent/DNA complexes are needed to achieve high transfection efficiency. The global charge of the complexes is determined by the PEI/DNA ratio (N/P ratio i.e. number of nitrogen residues (N) in the PEI per phosphate (P) of DNA). To obtain positively charged complexes, an N/P ratio of greater than 3 is needed.
The number of nitrogen residues available in the PEI depends on the molecular weight of the polymer, its structure (branched or linear), the deprotection of the protonable residues (deacylation) and the distribution of the fragment length (polydispersity) following hydrolysis.
The linear form of PEIpro™ and the manufacturing process developed by Polyplus-transfection ensure a high, stable and reproducible amount of protonable amines available for transfection while providing a truly deacylated molecule and an extremely lower polymer chain length variation.
The quality of PEIpro™ is continuously assessed during the manufacturing process with the appropriate control testing (Fig. 1). Further, systematic lot management and release testing is performed for each lot produced. To assess activity a standardized transfection assay is performed with a suspension-adapted HEK-293 cell line in animal-component free media to ensure the reliability and reproducibility of the transfection efficiency under conditions that are suitable for biomanufacturing.
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Fig. 1. PEIpro™ in-process and lot release quality controls. |
PEIpro™ is manufactured and formulated using a highly controlled production process (Fig. 1). Moreover, in order to meet or exceed current regulatory guidelines, PEIpro™ is released using advanced quality controls including a specification for a transfection efficiency that enables excellent lot-to-lot consistency (Fig. 2).
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Fig. 2.HEK-293 EBNA cells were seeded in synthetic media, incubated at 37°C, 8% CO2 with constant shaking and transfected with PEIpro™ following the standard protocol. Luciferase expression was assayed 48 h after transfection. |
PEIpro™ is free of component of animal-origin.
In addition, Polyplus-transfection is ISO 9001 Quality Management System accredited since 2002 and this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.
To ensure reliable production of recombinant proteins during the whole development process, any PEI transfection reagent selected should have the potential to meet cGMP guidelines from FDA, EMA, or ICH when the transfection reagent will be used to produce therapeutic recombinant proteins that are in late stage clinical development (ie, post Phase 1). PEIpro™ is tested to ensure complete sterility, absence of mycoplasma and very low level of bacterial endotoxins following pharmacopeia standards. In addition, on request, Polyplus-transfection can supply fully GMP compliant product delivering the highest quality level reagent and documentation for the production of therapeutic proteins or viral vectors.
Polyplus-transfection SA is the exclusive worldwide licensee of U.S. Patent No. 6,013,240, European Patent No. 0770140, and foreign equivalents, which cover transfection compositions having nucleic acids and a cationic polymer based on polyethylenimine (PEI), and use of this cationic polymer in nucleic acid transfection.
Polyplus sells branded PEI-based transfection reagents under its exclusive license, including the PEIpro™. For PEIpro™ transfection reagent sold by Polyplus and its authorized distributors, Polyplus hereby conveys license rights to the buyer to use the purchased transfection reagents, to the exclusion of human use, to perform stable or transient transfection for research and commercial purposes.
No rights to reverse engineer or resell are conveyed for any purchased transfection reagent. No license rights whatsoever are conveyed for any PEI-based molecules purchased from unauthorized sources for transfection purposes, and Polyplus reserves all rights and remedies relating to such unauthorized purchases.
jetPEI™-FluoR (red)
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Complexes formation is perfectly performed with NaCl solution. 0.5 ml of fluorescent jetPEI™ is sufficient to performed 250 transfections in 24-well plates. |
jetPEI™-FluoF (green)
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Complexes formation is perfectly performed with NaCl solution. 0.5 ml of fluorescent jetPEI™ is sufficient to performed 250 transfections in 24-well plates. |
PULSin™
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0.4 ml of PULSin™ protein delivery reagent is sufficient for 24 reactions in 6-well plates. For bulk quantities, please contact us. |
PROTEINS
PULSin™ was shown to deliver R-phycoerythrin, a fluorescent protein (240 kD) to the cytoplasm of up to 98% cells. As shown in Figure 1, the protein is evenly distributed in the cytoplasm and excluded from the nucleus due to its large size.
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Fig 1. PULSin™-mediated intracellular delivery of R-phycoerythrin to NIH-3T3 cells. R-phycoerythrin (1 µg) was complexed with 4 µl of PULSin™ for 15 min and added to NIH-3T3 cells in a 24-well plate. Live cells were observed by fluorescence microscopy after 16 h. |
ANTIBODIES
Antibodies were also successfully delivered to HeLa cells and able to recognize their target protein inside the cytoplasm.
For example, PULSin™ permits the delivery of FITC-labeled anti-alpha-tubulin to the cytoplasm of 85% HeLa cells (Fig.2).
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Fig 2. Delivery of a fluorescein-conjugated anti-alpha-tubulin antibody with PULSin™ to HeLa cells. |
Similarly, anti-giantin Alexa Fluor® 488 was delivered to the cytoplasm of 98% of live HeLa cells, labeling the Golgi apparatus (Fig.3).
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Fig 3. Golgi labeling (green) of HeLa cells 24 h after delivery of 1 µg Alexa Fluor® 488 anti-Giantin using PULSin™. |
Plasma membrane was stained with ConA-rhodamine. Cells were observed by confocal microscopy.
PEPTIDES
Peptides are biomolecules acting with high specificity at low concentrations. The delivery of substrate, inhibitor, modulator, or blocking peptides into cell allows protein function studies as well as the development of therapeutic approaches. PULSin™ was shown to successfully deliver Streptococcus TPE B epitope into HeLa cells (Fig. 4).
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Fig 4. Delivery of Pep-A (Streptococcus TPE B epitope, 16 aa), into HeLa cells. Complexes were formed with Pep-A (1 µg, lissamine-rhodamine derivative, Sigma) and PULSin™ (4 µl). Observation was carried out 16 h post-delivery. |
Other proteins, antibodies and peptides have also been delivered to cells using PULSin™ (Table 1).
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Table 1. Examples of proteins, antibodies and peptides delivered to cells using PULSin™ |
| PULSin™ will save you time and efforts compared to other techniques using viral transduction or chemical conjugation.PULSin™ reagent is ready-to-use and provided with a dilution buffer and a fluorescent control protein (R-phycoerythrin).
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PULSin™ was shown to deliver proteins and antibodies to a large variety of cell lines and primary cells (Table 2, Fig. 1-3).
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Table 2. Efficiency of R-phycoerythrin delivery using PULSin™ in selected cells. |
The comparison of PULSin™ with two other protein delivery reagents showed a higher efficiency of protein delivery (Fig. 5). Moreover, the amount of protein delivered per cell was higher with PULSin™ as measured for R-phycoerythrin protein and for FITC-alpha-tubulin antibody (Fig. 6).
Fig 5. Comparison of PULSin™ efficiency with two other protein delivery reagents. R-phycoerythrin (1 µg) was complexed with each reagent according to the manufacturer’s protocol. Complexes were added to HeLa cells and observed by fluorescence microscopy over 24 hours.
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Fig 6. FACS analysis of HeLa cells after delivery of R-phycoerythrin (upper graph) or FITC-anti-alpha-tubulin antibody (lower graph) with PULSin™ or with competitor reagents B and C.Data are presented as histograms of the mean intensity of fluorescence for each cell population. |
PULSin™ contains a proprietary cationic amphiphile molecule that forms non-covalent complexes with proteins and antibodies.
Complexes are internalized via anionic cell-adhesion receptors and are released into the cytoplasm where they disassemble. The process is non-toxic and delivers functional proteins.
Weill, C., Biri, S., and Erbacher, P. (2008b). Cationic lipid-mediated intracellular delivery of antibodies into live cells. Biotechniques 44, Pvii-Pxi.7
Weill, C., Biri, S., Adib, A., and Erbacher, P. (2008a). A practical approach for intracellular protein delivery. Cytotechnology 56, 41-48.
Transfection of Insect Cells
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1 ml of FectoFly™ is sufficient to perform 100 to 200 transfections in 6-well plates. For bulk sizes, Please contact us. |
By simple and rapid transient transfection method, FectoFly™ provides high transfection efficiency in insect cells and robust protein production over several days (Fig. 1).
FectoFly™ provides high transfection efficiency in most commonly used insect cells (Fig. 2) allowing efficient protein production.
FectoFly™ allows successful transfection of adherent and suspension insect cell cultures both in the latest generation of syntheticserum-free media (Fig.1, 3 – Sf21 and Sf9) and in the presence of serum (Fig.3 – S2). Transfection efficiency ranges routinely from 30% to 50% and cell viability is above 80%.
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Fig 3. GFP expression in Sf21, Sf9 cells grown in suspension, 72 h after transfection of pCMV-EGFP-Luc plasmid using FectoFly™. |
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FectoFly™ was compared to other commercially available reagents dedicated to the transfection of insect cells (Fig. 4). As shown for Sf9 cells, the highest protein expression levels were obtained with FectoFly™.
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Transfection of insect cells with FectoFly™ is straightforward.
Figure 5. FectoFly™ standard protocol.
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jetPRIME™
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1.5 ml is sufficient to perform ca.375 transfections in 6-well plates.Bulk quantities are available upon request.Please contact us. |
Superior transfection efficiencies ranging between 70 and 90% were obtained when using jetPRIME™ reagent versus the top competitor’s reagent for several commonly used cell lines. (Fig. 1).
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Fig. 1. Transfection efficiency assessed by FACS analysis in various cell lines 24 h following transfection in 24-well plates according to the manufacturer’s recommendation for competitor L2K and 0.5 µg plasmid, 1 µl reagent per well for jetPRIME™. |
Many other cell lines of various origins, as well as primary cells, are transfected with unusually high percentages (Table 1).
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Table 1. Transfection efficiency of various cell types using jetPRIME. The percentage of GFP-positive cells was determined by FACS analysis 24 h after transfection. |
jetPRIME™ is such a powerful in vitro transfection reagent that it only requires a small amount of reagent and plasmid DNA (Table. 2), making it very economical.
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Table 2. Amounts of reagent and DNA (jetPRIME™ and competitor) added per well in 6-well plate for transfection according to manufacturers’ recommendations. |
In addition to reducing costs, using less DNA also minimizes adverse cytotoxic effects triggered by transfection. Hence, jetPRIME™ is the reagent of choice for high transfection efficiency with excellent cell viability.
jetPRIME™ is extremely gentle to cells during transfection leading to increased cell viability (Fig. 2) and improved transfection results. Cells transfected with jetPRIME™ are healthy, while major cytotoxicity is observed with competitor.
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Fig. 2. Phase contrast microscopy of HeLa cells 24 h after transfections performed according to the manufacturer’s recommendations for each reagent. |
jetPRIME™ leads to over 90% knockdown of endogenous gene expression in a variety of cell lines. For example, jetPRIME™-mediated transfection of HeLa cells with 10 nM siRNA duplexes targeting endogenous lamin A/C in HeLa cells drastically reduces lamin A/C gene expression to barely detectable level (Fig. 3).
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Fig. 3. Endogenous lamin A/C silencing using jetPRIME™. HeLa cells were transfected with 10 nM of 21-mer lamin A/C siRNA. After 48 h, lamin A/C silencing was assessed by immunofluorescence microscopy using an antibody against lamin A/C. |
jetPRIME™ is well suited for DNA and siRNA cotransfection experiments. It shows highly efficient gene silencing in a variety of cell lines with very low toxicity. Over 90% silencing is achieved in adherent cells, using 10 nM siRNA (Fig. 4).
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Fig. 4. Exogenous luciferase gene silencing in several cell lines after DNA & siRNA cotransfection using jetPRIME™ performed with 400 ng pCMV-Luc and 10 nM of luciferase siRNA per well in 6-well plates. |
jetPRIME™ is an easy-to-use transfection reagent (Fig. 5):
• Fast and easy to scale up and down
• Compatible with serum and antibiotics
Fig.5. jetPRIME™ convenient protocol for DNA, siRNA and co-transfection of DNA and siRNA. |
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jetPEI™-HUVEC
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Complexes formation is perfectly performed with NaCl solution. 0.5 ml of jetPEI™-HUVEC is sufficient to perform 125 transfections in 24-well plates. |
The effect of serum concentration and incubation time on transfection efficiency was analyzed (Fig. 1). Transfection experiments performed in low (2%) or high (30%) concentrations of serum have shown that jetPEI™-HUVEC is most efficient when the cells are grown in low serum concentrations. Gene expression reaches a maximum for HUVEC after 4 h incubation with DNA/jetPEI™-HUVEC complexes.
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Figure 1. Effect of low (2%) versus high (30%) serum concentrations on transfection efficiency of jetPEI™-HUVEC. HUVEC cells were transfected for 2, 4 or 24 h with 2 µg of pCMVLuc plasmid DNA and 4 µl of jetPEI™-HUVEC in 24-well plates. Luciferase activity was measured 24 h after transfection. |
Compared to jetPEI™, our standard and versatile transfection reagent, jetPEI™-HUVEC provides significantly higher transfection efficiencies in HUVEC cells (Fig 2). Indeed, jetPEI™-HUVEC induces less cellular toxicity than jetPEI™. Using the optimal conditions for jetPEI™-HUVEC, transfection efficiencies of 50% have been reached as observed with a fluorescent protein reporter gene (Fig 3).
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Figure 2. Transfection efficiency and toxicity of jetPEI™ and jetPEI™-HUVEC in HUVEC. Cells were incubated for 4 h with transfection complexes in 2% serum medium. Luciferase activity was measured 24 h after transfection. |
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Figure 3. FACS analysis of HUVEC cells transfected with a plasmid encoding the EGFP fluorescent protein using jetPEI™-HUVEC. Transfection was performed in 24-well plate with 2 µg of plasmid and 4 µl of jetPEI™-HUVEC (N/P =5) and in 2% serum. Complexes were incubated for 4 h with the cells. FACS analysis to quantify EGFP expression was performed 24 h after transfection. |
jetPEI™-HUVEC compares particularly favorably to other non-viral transfection reagents whether cationic lipids or polyamine. jetPEI™-HUVEC compares particularly favorably to other non-viral transfection reagents whether cationic lipids or polyamine (Fig. 4).
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Figure 4. Comparison of transfection efficiency of jetPEI™-HUVEC and other commercially available reagents. Cells were transfected using optimal conditions established for each reagent. |
The jetPEI™-HUVEC protocol is as simple as that of jetPEI™: Mix the DNA with the reagent to form complexes and simply add the mixture to the cells. jetPEI™-HUVEC is compatible with serum and antibiotics, thus eliminating the need for media changes. Protein expression is determined 24 h to 72 h post-transfection.
