Overview

Specifications

Reagent

in vivo-jetRNA®+ transfection reagent

Molecule delivered

mRNA

Applications

Vaccination / immunization
Cancer therapy
in vivo functional studies (Gene expression, CRISPR genome editing)

Targeted organs

All organs (depending on the injection route)

Injection routes

Various administration routes (systemic or local)
Intravenous / intramuscular / intraperitoneal and retro-orbital injections have been successfully tested

Number of injections in mice

1 mL is sufficient to perform at least 50 intravenous injections or 100 intramuscular injections in mice

Storage

Store in vivo-jetRNA®+ at 5 °C ± 3°C. Do not freeze.
Expiry date is indicated in the certificate of analysis and on the product.

Provided with

mRNA buffer


Summary

in vivo transfection reagents are the most powerful alternative to viral vectors for nucleic acid delivery. They are easy to use, cost-effective and considered as safe and efficient vehicles for RNA delivery. mRNA transfection is rapidly emerging as a promising method for nucleic acid-based therapy and offers an attractive substitute to plasmid DNA. Non-viral mRNA delivery methods have already proven their efficiency in vaccination through antigen presenting cells modification and in anti-cancer therapy by directly targeting malignant cells. The intrinsic advantage of mRNA-based immunotherapy relies on the self-adjuvant activity of mRNA and the fact that small amounts of encoded antigen are sufficient to obtain robust immune response.

in vivo-jetRNA®+ is a lipid-based transfection reagent composed of preformed liposomes specifically developed to deliver mRNA in vivo. This reagent can be used to target unique or multiple organs, by using systemic injection routes, in various animal models (mice, rat, etc.). mRNA delivery using in vivo-jetRNA®+ is user-friendly, with a simple 2-step protocol and can be used for vaccination purposes, anti-cancer studies, genome editing using CRISPR/Cas9 method or protein replacement.

Webinar: Next generation lipid-based delivery for mRNA therapeutics: in vivo-jetRNA®+

Ordering information

Reference Number Volume of in vivo-jetRNA®+ Volume of mRNA Buffer
101000122 1 mL 60 mL

Bulk quantities are available upon request. Please contact us.

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Description

Efficient: 100% mRNA encapsulation leading to comparable LNP delivery results

in vivo-jetRNA®+ is the reagent of choice to deliver mRNA to various organs. This is attributable to its intrinsic properties: in vivo-jetRNA®+ protects its payload against ubiquitous endonucleases, prevent non-specific interactions with proteins and promote efficient cell entry. 

in vivo-jetRNA®+ shows an amazing capacity to encapsulate 100% of the mRNA available (Fig 1 A/) leading to an efficient gene expression that is comparable to lipid nanoparticles currently used as a gold standard for vaccination purpose (Fig 1 B/).

Fig1. in vivo-jetRNA®+ leads to 100% mRNA encapsulation leading to efficient mRNA delivery. A/ mRNA alone (lane 1) and mRNA fully encapsulated using in vivo-jetRNA®+ (lane 2) were analyzed by gel electrophoresis (agarose gel). B/ mRNA encoding Luciferase was injected into mice using in vivo-jetRNA®+ or Dlin-MC3-DMA LNP through intravenous injection. Liposomes were formed using 10 µg of mRNA with an mRNA/in vivo-jetRNA®+ ratio of 1:2 (µgmRNA:µLreagent) in mRNA Buffer. Luciferase expression was assessed 24 h post-injection.​

Ready-to-use: Liposome-based reagent that does not require any formulation equipment

Lipid nanoparticles (LNP) are a gold standard to deliver mRNA but it requires months of work, dedicated equipment and consumables to reach the optimal and efficient formulation. With in vivo-jetRNA®+, there is no need to spend time and budget to reach the perfect formulation. in vivo-jetRNA®+ has been carefully optimized by Polyplus and can be used with a simple and straightforward protocol without any formulation step.  

in vivo-jetRNA®+/mRNA liposome solution is prepared in two steps and ready to be injected to the animal in 15 minutes: a gain of time with minimal effort (Fig. 2). Contrary to LNP, in vivo-jetRNA®+ is a ready to use formulation that do not require any formulation equipment.

Fig 2. in vivo-jetRNA®+ simplified protocol. This two-step protocol is suitable with direct injection of in vivo-jetRNA®+/mRNA liposomes through any systemic or local administration route. Contrary to LNP, in vivo-jetRNA®+ is a ready to use formulation that does not require any formulation equipment.​

Our friendly scientific support team use their scientific expertise to provide protocols tailored to meet your needs.  

Contact Scientific Support

Stable: Liposomes are stable using low/high mRNA concentration 

In addition of the capacity to encapsulate mRNA, in vivo-jetRNA®+ excels in stability when it is formulated with mRNA. Liposome’s sizes are stable up to 1 month when incubated at room temperature or 4°C using in vivo-jetRNA®+ (Fig 3. A/). The mRNA concentration will influence the liposome’s size but in vivo-jetRNA®+ shows a stable liposome size at low or high mRNA concentration (Fig 3. B/). 

This makes in vivo-jetRNA®+ the reagent of choice for vaccine or mRNA cancer therapy. 

Fig 3. Liposomes (including in vivo-jetRNA®+ & mRNA) are very stable over the time using low to high mRNA concentration. A/ Size of liposomes with in vivo-jetRNA®+ at 50 μg/ml after 15 min, 1 week, 2 weeks, 3 weeks and 4 weeks of complexation stored at room temperature (RT) or 4°C were measured with the dynamic light scattering (DLS). B/ Size of liposomes with in vivo-jetRNA®+ at 50, 100, 200 or 300 μg of mRNA/ml with a small mRNA (1929b) were measured by DLS.

Universal: Target any organ/tissue using various administration routes 

Choice of the administration route leads to different mRNA biodistribution. in vivo-jetRNA®+-mediated mRNA delivery leads to gene expression in numerous organs, through intraperitoneal injection, especially in the spleen and nodes that have a major role in the immune response but also in other organs such as lung, liver, pancreas and uterus (Fig. 4).  

Being a non-viral delivery reagent, in vivo-jetRNA®+ can be safely administered in animals. By avoiding conventional vaccine approaches, it allows to generate a new class of vaccines that rely on direct gene transfer. This concept has several advantages – it is safer for the recipients, faster and cheaper to produce compared to viral vector vaccines or peptide loaded APC (Antigen Presenting Cells), and it has an immense potential to tackle many unmet medical needs.

Fig 4. in vivo-jetRNA®+ leads to efficient mRNA delivery in different organs depending on the administration route. mRNA encoding Luciferase was injected into mice using in vivo-jetRNA®+ through different administration routes A/ intraperitoneal (IP) and B/ intramuscular (IM). Liposomes were formed with a mRNA/in vivo-jetRNA®+ ratio of 1:2 (µgmRNA:µLreagent) in mRNA Buffer using either 20 µg mRNA for intraperitoneal (IP) injection or 5 µg mRNA for intramuscular (IM) injection. Luciferase expression was assessed 24 h post-injection.

Safe: Maintain healthy animals and organs

in vivo-jetRNA®+ is the best choice in terms of safety. There are no secondary effects after injection, all animals remain healthy and no animal pain has been encountered. After administration of mRNA – in vivo-jetRNA®+ liposomes by any administration route, targeted organ remains phenotypically intact and no tissue damage is observed. 

Furthermore, using in vivo-jetRNA®+ triggers no pro-inflammatory cytokine expression (Fig 5.)

Fig. 5 in vivo-jetRNA®+ triggers no pro-inflammatory cytokine expression. mRNA liposomes were formed in 200 µL of mRNA Buffer using 10 or 20 µg of mRNA encoding Luciferase at a mRNA/in vivo-jetRNA®+ ratio of 1:2 (µgmRNA:µLreagent) and injected through intravenous injection (retro-orbital injection). 2 to 24 hours after injection, blood was collected and the level of IL-6, IL-12, GM-CSF, IFN-gamma and TNF-alpha was measured by ELISA (IL-6) or MACSPlex kits. As a positive control, LPS (200 µg) was administered into mice.

Applications

Immunization/vaccination

Immunization or vaccination is a process of introducing novel antigens to the immune system and enabling their recognition, leading to a specific immune response later on. Instead of presenting the whole inactivated pathogens or their partial forms as proteins/peptides to the immune system, mRNA coding for a relevant antigen complexed with in vivo-jetRNA®+ can be administered to the organism. This mRNA vaccine leads to generation of the appropriate antigen in the organism which elicit the subsequent immune response. 

in vivo functional studies

in vivo-jetRNA®+ is ideal to study gene function in vivo and provides the easiest method for the corroboration of in vitro study. in vivo-jetRNA®+ is especially suited for gene expression in cells that are usually difficult to transfect, such as slow diving cells or cells that have developed specific mechanisms to protect their genome. 

in vivo gene editing using CRISPR/Cas9 system

The latest gene editing tool, CRISPR/Cas9 system, has proven to be an elegant solution for gene therapy. CRISPR/Cas9 in combination with in vivo-jetRNA®+ provides both precise delivery and effectiveness, giving rise to promising anti-cancer therapeutics, among other applications.

Quality

Each lot of in vivo-jetRNA®+ is tested in-house with a proprietary test following standard protocol. A sterility test is assessed for each lot. Every batch will be provided with Certificate of Analysis. 

Polyplus-transfection® is ISO 9001 Quality Management System accredited since 2002; this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.

Protocol

In order to download a product protocol or a certificate of analysis, please create an account on Polyplus-transfection® Portal.

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