Viromer® in vivo mRNA Systemic Kit

IN VIVO mRNA transfection reagent

The Viromer® IN VIVO mRNA transfection reagent is a lyophilized powder containing Viromer® nanoparticles. It only needs to be rehydrated with the diluted target mRNA to form active particles.

The Systemic Kit contains:

  • 6 vials of lyophilized Viromer® (30rxns)
  • 2 vials of lyophilized Positive Controls (2rxns, Luc encoding mRNA complexed to Viromer®)
1.250,00
Article No.:
Vivo-01mRNA-02

General Description

Lipocalyx Viromer® IN VIVO mRNA transfection reagent facilitates effective systemic delivery and protein expression in animals. Viromer® IN VIVO reagents are loaded with an mRNA provided by the user by simply adding an mRNA solution to the lyophilized product. Systemic injection into the tail vein of a mouse or another animal results in rapid protein expression in liver and spleen. For luciferase, substantial activity as early as two hours upon injection was detected and the signal could be tracked for up to 44 hours.


Systemic use - exclusive spleen targeting

A systemic (intravenous) injection of 10µg mRNA or less per mouse results in a specific targeting of the spleen. Delivery of mRNA results in the local production of the encoded protein and an expression of 150ng luciferase per gram of spleen tissue 6h post-injection was reported. Proteins having an export sequence or signal peptide such as EPO are excreted into the circulation; serum levels of 60 ng/mL upon a single injection of 10µg mRNA as early as 2h after the injection have been measured.

 

picture of a luciferase signal in the spleen of a mouse transfected with the Viromer® in vivo mRNA transfection reagent from the dorsal side          picture of a luciferase signal in the spleen of a mouse transfected with the Viromer® in vivo mRNA transfection reagent from the ventral side

Figure 1: Luciferase signal 6h after a single dose of 10µg mRNA i.v.

 


Systemic use - combined spleen and liver targeting

Intravenous injections of 20 to 40µg mRNA per mouse targets the liver in addition to spleen. Typical levels of expression are 50 and 400ng luciferase per gram of liver or spleen tissue, respectively. For EPO, a secreted protein, the injection of 30µg mRNA complexed to Viromer gave serum concentrations of 160 ng/mL at 2h after the injection.

 

picture of a luciferase signal in the spleen of a mouse transfected with the Viromer® in vivo mRNA transfection reagent from the dorsal side          picture of a luciferase signal in the spleen of a mouse transfected with the Viromer® in vivo mRNA transfection reagent from the ventral side

Figure 2: Mice received a single dose of 40µg luciferase encoding mRNA:Viromer complexes i.v. (6h after application)

 


Applications

Given the specific targeting to spleen and the very high performance of Viromer with macrophages, the technology is ideally for immune-related applications such as:

  • vaccination
  • immunostimulation
  • anti-infective approaches
  • testing of anti-inflammatory compounds
  • testing of immunosuppressive compounds
  • Related cargoes are: OVA (model antigen)

Apart from targeted approaches to the immune system, the Viromer IN VIVO for the first time facilitates the effective expression of a variety of proteins from mRNA.

  • enzyme replacement studies
  • expression of fluorescent proteins
  • gene editing
  • Related Cargoes are: LUC (cytosolic, facilitates in vivo tissue distribution studies), GFP (cytosolic and cell specific), EPO (secreted)

Key Specifications

The reconstituted Viromer IN VIVO has the following specifications:

  • Particle size                       250nm
  • Surface charge                 -10 ….+10mV (close to neutral)
  • % of free mRNA              <10%

Features and Benefits

  • High transfection efficiency due to an active escape of Viromer® complexes from the endosome.
  • Great safety because Viromer complexes are charged close to neutral (-10 ….+10mV).
  • We have noticed a striking difference in the transfection of mRNA when compared to DNA coding for the same protein. While DNA works well in many cultured cells, there is a much better expression for mRNA in hepatocytes and macrophages. The latter are working almost exclusively with mRNA. We hypothesize that innate immunity rather than the actual transfection is behind this difference and cytosolic sensors such cGAS/STING or AIM2 have been described. In practical terms, we highly recommend the use of mRNA for transgene expression in animal studies.
  • Easy and fast transfection with consistent results using a lyophilized transfection reagent ascribed to a straightforward protocol.

Viromer IN VIVO mRNA transfection protocol systemic kit


Notes

  • For research use only! Not be used in humans or for diagnostics. Ethical regulations apply but may vary between institutions and you might be required to obtain permission for your experiments.
  • Sources of mRNA: Lipocalyx recommends using mRNA from Trilink, Inc.
  • store dry at 2-8°C
  • The technology is available for pharmaceutical development, please contact info@lipocalyx.de for further discussion.