Messenger RNA (mRNA)-based vaccines have shown great potential to activate antitumor immune response for cancer therapy. Numerous unsolved technical difficulties in successful in vivo delivery of in vitro transcribed (IVT) mRNA, such as transfection efficiency, have greatly hindered the applications of IVT-mRNA in clinical development. As one of the lipid-based vectors, lipoplexes have recently been the subject of intensive investigations to understand the parameters governing the efficiency of transfection.
Equipped with world-leading technology platforms and professional scientific staff in the development of the drug delivery system, Creative Biolabs is committed to providing high-quality lipoplex-based delivery systems to efficiently facilitate targeted in vivo delivery of IVT-mRNA in safe manner.
mRNA is frequently encapsulated with positively charged liposomes to form lipoplexes. Lipoplexes are self-assembling nanosystems and their formation entails a multistep mechanism as revealed by detailed studies employing monolayers, atomic force microscopy and cryo-electron microscopy. In a millisecond first step, electrostatic interactions between phosphate and the positively charged amine headgroup of the cationic lipid occur. The cellular uptake is mainly mediated by either lipoplex endocytosis or endocytosis-like mechanisms. Moreover, lipoplex delivery systems have substantial advantages like easy synthesis, low batch variability, scalability, and biocompatibility. It has also been noted that drug delivery systems in the nano-range enhance the pharmacokinetic availability of encapsulated drugs.
IVT-mRNA has been applied as a therapeutic molecule in the field of cancer therapy and biomedical research studies. mRNA-based therapy has shown several advantages but its further application is largely restricted by delivery method.
Researchers have developed a liposome-protamine lipoplex (CLPP) to deliver mRNA encoding survivin-T34A gene, resulting in a novel core-shell structured nanoparticle formulation (CLPP/mSur-T34A). This CLPP/mSur-T34A formulation displayed high mRNA transfecting and expression efficiency on C26 tumor cells through lipid rafts-mediated endocytosis. CLPP/mSur-T34A mRNA formulation showed obvious therapeutic effects on various models of C26 colon cancer both in vitro and in vivo. The results indicated the high delivery capacity of lipoplex and further suggested that CLPP/mSur-T34A mRNA formulation can be a potential candidate for colon cancer therapy.
Another team reported a lipoplex-based mRNA nanovaccine which can greatly improve mRNA delivery in antigen-presenting cells, activate T cell response, and induce robust antitumor immunity. This lipoplex-based formulation is composed by cationic polymer/lipid(poly(amidoamine)/alkylene oxide), DSPE-PEG2000, and epitope-encoding mRNA. With appropriate ratio, the optimized nanovaccine demonstrated effective mRNA delivery in dendritic cells, and strongly active antigen-specific immune response in vitro and in vivo.
Fig.1 Negatively charged IVT-mRNA spontaneously interact with cationic lipids to form lipoplexes.1
At Creative Biolabs, liposomes are usually composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-distearoyl-sn-glycero3-phosphoethanol-amine-N-[biotinyl(polyethylene glycol)-2000] (DSPE-PEG-2000-biotin). With years of experience in nanotechnology and material science, our scientists are more than happy to share our cutting-edge technologies and experience in the development of lipoplex-based delivery system for the transfection of therapeutic mRNA. Moreover, we also provide a comprehensive analysis of mRNA-lipoplexes, including transfection efficiency, duration of protein expression, immunogenicity, hemocompatibility, and storage stability.
To assist our clients in the development of lipoplex-based delivery systems, Creative Biolabs is proud to offer the most comprehensive services. For more detailed information, please feel free to contact us or directly send us an inquiry.
Inquire About Our ServicesA: Lipoplex-based delivery systems are complexes formed by the interaction between cationic lipids and negatively charged nucleic acids (such as DNA or RNA). These systems facilitate the delivery of genetic material into cells by promoting cellular uptake and protecting the nucleic acids from degradation.
A: Creative Biolabs offers comprehensive services for the development and optimization of lipoplex-based delivery systems. These services include formulating and characterizing lipoplexes, optimizing transfection efficiency, assessing stability and biocompatibility, and conducting in vitro and in vivo delivery studies to ensure effective and targeted delivery of nucleic acids.
A: Creative Biolabs employs advanced formulation techniques to optimize the ratio of cationic lipids and nucleic acids, particle size, surface charge, and other parameters to enhance delivery efficiency and minimize toxicity. They also use high-throughput screening and detailed biophysical characterization to identify the most effective lipoplex formulations.
A: Lipoplex-based delivery systems offer several benefits, including high encapsulation efficiency, protection of nucleic acids from degradation, ease of preparation, and the ability to deliver a variety of genetic materials. These systems also facilitate endosomal escape, ensuring that the nucleic acids reach the cytoplasm or nucleus to exert their desired biological effects.
A: Yes, Creative Biolabs provides custom lipoplex formulation services tailored to specific research or therapeutic goals. We can adjust the lipid composition, nucleic acid payload, targeting ligands, and other components to meet the unique requirements of different applications, including gene therapy, RNA interference, and vaccine development.
A: Creative Biolabs conducts comprehensive analyses of mRNA-lipoplexes, focusing on transfection efficiency, duration of protein expression, immunogenicity, hemocompatibility, and storage stability to ensure high efficiency and effectiveness.
This study focuses on the efficiency of mRNA delivery using lipoplexes prepared via a modified ethanol injection (MEI) method. Researchers evaluated 18 mRNA lipoplexes composed of different cationic lipids, neutral helper lipids, and PEG-Cholesterol. The mRNA lipoplexes containing DC-1-16 or TC-1-12 lipids exhibited high protein expression both in vitro and in vivo. Specifically, the DC-1-16, DOPE, and PEG-Chol composition showed significant protein expression in the lungs and spleen of mice, also inducing high antigen-specific IgG1 levels upon immunization. These findings suggest that the MEI method enhances mRNA transfection efficiency, making it a promising approach for mRNA-based therapies and vaccines.
Fig.2 mRNA biodistribution and protein expression in mice after IM injection of mRNA lipoplexes.2
| Cat. No | Product Name | Promoter |
|---|---|---|
| CAT#: GTVCR-WQ001MR | IVTScrip™ pT7-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ002MR | IVTScrip™ pT7-VEE-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ003MR | IVTScrip™ pT7-VEE-mRNA-FLuc Vector | T7 |
| CAT#: GTVCR-WQ87MR | IVTScrip™ pT7-VEE-mRNA-Anti-SELP, 42-89-glycoprotein Vector | T7 |
| Cat. No | Product Name | Type |
|---|---|---|
| CAT#: GTTS-WQ001MR) | IVTScrip™ mRNA-EGFP (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| CAT#: GTTS-WK18036MR | IVTScrip™ mRNA-Human AIMP2, (Cap 1, Pseudo-UTP, 120 nt-poly(A)) | Enzyme mRNA |
| (CAT#: GTTS-WQ004MR) | IVTScrip™ mRNA-Fluc (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| (CAT#: GTTS-WQ009MR) | IVTScrip™ mRNA-β gal (Cap 1, 30 nt-poly(A)) | Reporter Gene |
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