High-Efficiency Transfection Without Compromise

Achieve Robust mRNA and DNA Delivery Across Challenging Cell Models with Low Toxicity, High Reproducibility, and a Simple Non-viral Workflow

RJH transfection reagents offer a powerful alternative to traditional liposome based and viral delivery systems by combining high efficiency with an exceptionally favorable safety and handling profile. Unlike liposomal reagents, which can trigger cytotoxicity, membrane destabilization, or immune activation, RJH formulations are designed to be milder on cells, enabling robust expression with minimal impact on viability and phenotype.

In contrast to viral vectors, often labor intensive, costly, and subject to biosafety constraints, RJH reagents provide a simple, non viral approach that avoids genomic integration risks while still achieving strong delivery performance across a broad range of primary cells and hard to transfect models.

This balance of efficiency, reproducibility, and low cellular stress makes RJH reagents a versatile and reliable choice for researchers seeking consistent results without the complexity or safety concerns associated with liposomes and viral systems.

mRNA Delivery

It is a cationic lipopolymer transfection reagent optimized for the delivery of synthetic mRNA into both attachment and suspensions cells. It is a non-integrating carrier, so that the genetic make-up of host cells is not altered after the treatment. The reagent facilitates formation of stable nanoparticles, with sizes ranging between 100 to 200 nm, by forming robust interactions with mRNA molecules.

Figure 1: Transfection of GFP-mRNA with mRNA-Fect in attachment-dependent MCF-7, MDA-MB-436 and MDA-MB-231 cells. The expression levels were quantitated by flow cytometry 72 hours after transfection and summarized as the percentage of cells positive for GFP. For comparison, a leading lipofection reagent was used according to the manufacturer’s instructions, with mRNA-Fect demonstrating superior performance. 

It is an improved version of mRNA-Fect after extensive testing of amphiphilic polymer libraries.

Figure 2: Transfection of GFP-mRNA with mRNA-Fect ULTRA in suspension-growing MOLM-13, MV4-11, and Jurkat cells. GFP expression levels were quantified by flow cytometry 72 hours after transfection and summarized as the mean fluorescence intensity (MFI) of all cells. For comparison, Lipofectamine MessengerMAX was used according to the manufacturer’s instructions, with mRNA-Fect ULTRA demonstrating superior performance.

mRNA-Fect kit and mRNA-Fect ULTRA kit are the solution!

These kits include Trans-Booster, a synthetic polymer that is specifically tailored to enhance the transfection efficiency.

Figure 3: Transfecting hBMSC (right) and cbMSC (Left) cells with mRNA-Fect Kit. An mRNA-GFP was used to assess the efficiency of mRNA expression. Typical GFP expression levels were analyzed through flow cytometry. For comparison, a leading lipofection reagent was used according to the manufacturer’s instructions, with mRNA-Fect Kit demonstrating superior performance.

Figure 4: Neuro 2A cells transfected with a GFP-expressing mRNA using mRNA-Fect with and without Trans-Booster. The Trans-Booster enhances transfection efficiencies with mRNA in N2a cells.

It is a high concentration formulation of mRNA-Fect to allow high amounts of mRNA delivery typical of preclinical models.

Figure 5: GFP expression in tibialis anterior muscles and Luciferase expression in different vital organs of NCG mice. NT (No treatment): Injection with RPMI. mRNA-Fect In Vivo/mGFP or Luc-mRNA: mGFP injection with mRNA-Fect In Vivo formulation as part of the mRNA-Fect In Vivo. The animals were analyzed after 2 days of injection for the gene expression. A robust GFP or Luc expression was evident in the animals treated with reporter gene complexed using the mRNA-Fect.

If expression is still low after testing different conditions, consider using the mRNA Fect In Vivo Kit with the included Trans Booster to enhance transfection efficiency.

DNA Delivery

ALL-Fect and Prime-Fect are synthetic amphiphilic polymers, highly effective delivery agents tailored for plasmid DNA delivery.

Figure 6: TRAIL pDNA delivery with ALL-Fect In Vivo Kit. Human breast cancer xenografts were grown in NOD-SCID mouse and a TRAIL expressing plasmid DNA was administered subcutaneously with ALL-Fect In Vivo Kit in the vicinity of tumors. Left graph indicates the relative tumor growth (normalized with tumor volume on day 1) as a function of time. Middle graph indicates TRAIL protein expression in recovered xenografts (day 15) after homogenization. Right graph indicated mRNA levels of TRAIL protein in recovered xenografts. ALL-Fect In Vivo Kit was able to sustain the TRAIL expression to obtain a function outcome in retarding the tumor growth.

Benefits of RJH Transfection Reagents

• High transfection efficiency: Provides 2 to 7-fold higher efficacy in the presence of serum compared with lipofection reagents
• Simple Protocol: No need to change tissue culture medium during transfection
• Lower Toxicity: Less toxic compared to lipofection reagents, leading to minimal alteration of cell physiology
• No immunogenicity: No genome integration.

Simplify your workflow with RJH Transfection Reagents

Figure 7: Workflow set-up for experiments using transfection reagents from RJH

• References

  Article content created by Tebubio using courtesy materials provided by RJH Biosciences.

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