In vivo Delivery Reagents

In-vivo-jetPEI® is a polymer based reagent, successfully used to deliver DNA and siRNA for research purposes such as functional studies, RNAi in vivo studies, gene therapy or genetic vaccination.

In addition, in vivo-jetPEI® is currently used in clinical trials as a delivery vehicle for therapeutic goals, including viral diseases and cancer.

  • Successful in vivo delivery of DNA, siRNA and oligonucleotides
  • Currently used in Phase I & II clinical trials
  • Multiple modes of administration in many species
  • No detectable inflammatory response

Ordering information

Catalog number Amount of reagent Amount of glucos solution
201-10G 0.1 ml 10 ml
201-50G 0.5 ml 2 x 10 ml

0.1ml of in vivo-jetPEI® is sufficient to perform up to 20 intravenous injections in mouse with 50 µg of DNA. Formation of complexes with the provided glucose solution produces nanoparticules adapted to in vivo injections. Bulk quantities and GMP grade are available upon request. Please contact us.

Successful in vivo delivery of DNA, siRNA and oligonucleotides

in vivo-jetPEI® is the reagent of choice to deliver various types of nucleic acids, such as DNA, siRNA and oligonucleotides in order to mediate gene expression in various tissues upon in vivo administration as shown by in vivo imaging (Fig. 1).  in vivo-jetPEI® is so versatile that it can be used for different types of nucleic acid such as siRNA, shRNA plasmids, miRNA, oligonucleotides.

in vivo mouse 1






Figure 1. Luciferase expression following systemic delivery of a pCMVLuc plasmid using in vivo-jetPEI®. Bioluminescent imaging of luciferase expression in living Nude mouse using IVIS 100 camera (Caliper-PerkinElmer) 24 h after gene delivery. pCMVLuc (50 µg) was complexed with in vivo-jetPEI® in 400 µl of 5% glucose solution and injected into the tail vein.

The success of nucleic acid therapy relies on the ability to efficiently deliver the appropriate therapeutic materials into the target tissue or cells with low toxicity and limited immune response.

Hence, to explore the potential of in vivo-jetPEI® to deliver both plasmid DNA and siRNA in animals, we first administrated systemically a plasmid expressing the luciferase gene which resulted in maximal luciferase expression in the lungs (Fig. 2).

In a second experiment, siRNA matching the luciferase sequence was mixed with the plasmid and complexed with in vivo-jetPEI®prior to systemic injection (Fig. 2). A scramble siRNA was used as a negative control (Fig. 2). Systemic administration of a siRNA directed against luciferase showed a mean silencing superior to 90% as compared to the negative control Hence, specific inhibition of protein synthesis can be achieved by in vivo-jetPEI® mediated siRNA delivery in animal models.

 in vivo mouse 2







in vivo mouse 3 (1)

Figure 2. RNA interference in the lung with in vivo-jetPEI®. pCMVLuc (40 µg) was co-transfected with a control siRNA (upper) or with 10 µg of specific anti-Luc siRNA (lower). The complexes were injected into the tail vein of nude mice. Luciferase gene expression was monitored in living mice 24 h later by bioluminescence imaging using a cooled CCD camera.

Currently used in Phase I and II clinical trials

in vivo-jetPEI® was selected as a delivery vector at early stages in several drug development programs, due to its safety and high delivery efficiency. There are currently several ongoing phase I and II clinical trials for cancer therapies, heart disease, brain injury and HIV immune gene therapy using in vivo-jetPEI® (Fig. 3). Indeed, in vivo-jetPEI® is available at cGMP grade for use in humans as well as in preclinical grade for toxicology and biodistribution studies. Below are referenced the publications of groups currently involved in a clinical/preclinical program using in vivo-jetPEI®.

As an example of the use of Polyplus’ transfection® reagents in a cell therapy trial please visit the OHRI in Canada.

Multiple modes of administration in many species

The stability of in vivo-jetPEI® nucleic acid complexes allows the use of numerous routes of administration as shown in Figure 4. Depending on the route of administration, in vivo-jetPEI®-mediated gene expression was also observed in the brain, liver, pancreas, spleen, kidney, heart, bladder, skin, retina, artery, etc. More literature references are available on the Polyplus Product citations online database.

Routes and target organs

Upon intravenous injection, in vivo-jetPEI®-mediated DNA delivery leads to gene expression in the lung (Fig. 1 and Fig. 2) liver, pancreas, spleen, kidney, heart, bladder and artery. In addition, in vivo-jetPEI® is also well-adapted for local delivery such as application onto the skin, intratumoral, intracerebral or intra-articular injections (see Fig. 4).

 in vivo mouse 4


Fig. 4.

Animal models

in vivo-jetPEI® was successfully used to deliver nucleic acids to a wide range of species including mouse, rat, guinea pig, duck, rabbit, monkey, goat, sheep, chicken, quail, hamster, cow, tadpole, shrimp and fish. As a result, the experimental set up can be adapted to most species. Our technical support team will be pleased to help you adapt the protocol to your animal model, please contact us to find out more

Kinetics of transgene expression

Upon systemic delivery, nucleic acids are quickly delivered into organs and expressed in cells. The time course of luciferase expression in lungs after intravenous injection of a luciferase expressing plasmid complexed with in vivo-jetPEI® was investigated using Caliper-PerkinElmer bioimaging system (Fig. 5A) and luciferase expression assay on lung extracts (Fig. 5B). Expression of luciferase can be detected as early as 12 h post-injection, and is maintained for at least 72 h post-injection. Duration of transgene expression can be increased depending on the protein stability and half-life. In particular, the use of plasmids lacking CpG motives avoids silencing of gene expression over time and allows a longer expression of the transgene.

 in vivo mouse 5




in vivo fig 5

Figure 5: Kinetics of luciferase expression into lungs. pCMVLuc plasmid (40 µg) complexed with in vivo-jetPEI® was injected intravenously. (A) Mice were imaged 12, 24, 48, 72 and 96 h after injection using IVIS 100 bioimaging system (Caliper-PerkinElmer). (B) Lungs were extracted at the same time points and luciferase expression was measured using a standard assay on lung extract. 


No detectable inflammatory response

Viral vectors can elicit an immune response following injection; however minimizing side effects is crucial for in vivo research experiments and subsequent clinical applications. Non-viral vectors such as in vivo-jetPEI® offer a reliable and safe alternative. Following in vivo-jetPEI® systemic delivery of siRNA, there is no induction of major pro-inflammatory cytokines such as TNF-alpha, IL-6 and IL-12/IL-23 (Fig. 6). In addition, no increase in sera levels of hepatic enzymes are detected 24 h after complex injection, suggesting that in vivo-jetPEI® does not trigger hepatotoxicity. Hence, in vivo-jetPEI® does not induce any significant inflammatory response after systemic injection of siRNA (Fig. 6) and DNA (data not shown), making it the reagent of choice for safe in vivo nucleic acid delivery experiments (Bonnet et al. (2008), Pharm Res, 25:2972).

in vivo fig 6


Fig. 6. Serum concentration of TNF-α, IL12/IL23, IFNγ and IL6 following intravenous nucleic acid delivery using in vivo-jetPEI® (N/P=8) respectively 1h, 6h, 12h and 6 h after delivery. The negative control consists of an IV injection of 5% glucose; the positive control an IP injection of 50 μg E.ColiLPS. 40 ug siRNA were delivered with or without in vivo-jetPEI®.

Reproducible results

Gene delivery using in vivo-jetPEI® is reliable providing reproducible data (Fig. 7) with limited toxicity in contrast to other non-viral delivery methods. The reproducibility is linked to the unique properties of in vivo-jetPEI®. In 5% glucose, in vivo-jetPEI® condenses nucleic acids into stable nanoparticles of ca. 50 nm in diameter. As a result, aggregation of blood cells is reduced compared to other reagents (Kircheis et al. (2001), Gene Ther 8: 28), thereby preventing a restriction in diffusion within tissues, erythrocyte aggregation and microembolia.

in vivo fig 7



Fig. 7: Systemic delivery using in vivo-jetPEI®. pCMVLuc (50 µg) was complexed with in vivo-jetPEI® in 5% glucose and injected retro-orbitally. After 24 h, luciferase gene expression was assessed in the lung (n=8). Viability of mice was 100%.

A very easy to handle protocol

in vivo fig 9

Fig. 9: The protocol of in vivo-jetPEI® is very easy to use It consists in mixing the nucleic acid and the reagent in 5% glucose and then injecting the complexes into the animal. This protocol is fast and suitable for numerous administration route.

STICKY siRNA™ (ssiRNA): a novel approach

In order to improve siRNA delivery in vivo, Polyplus transfection has developed a novel type of siRNA. By including longer overhangs within the siRNA, we have generated STICKY siRNA™ (ssiRNA) that are able to form concatemers in the presence of in vivo-jetPEI®, thereby mimicking the structure of DNA and thus enhancing siRNA transfer. This was shown to improve in vitro siRNA delivery and gene silencing efficiency (Bolcato-Bellemin (2007), PNAS 104:16050). We have also shown that intraperitoneal injection of STICKY siRNA™ cyclinB1 in mice using in vivo-jetPEI® in a PC-3 tumor model, lead to an inhibition of tumor metastasis compared to mismatch and control. Moreover, mice survival is increased dramatically with STICKY siRNA™ delivered by in vivo-jetPEI®.