RJH Biosciences Inc.

RJH Biosciences Inc.Homepage
RJH Biosciences is a biotechnology company based out of Edmonton, Alberta, Canada. We develop novel transfection reagents and delivery systems that transport different types of nucleic acids to a range of human cells and cell lines. We develop value-added products in two commercial segments, as transfection reagents for biomedical R&D enterprise and as nucleic acid delivery vehicles for preclinical and clinical applications. The same transfection reagents are used in our own R&D projects that focus on blood cancers and modification of immune cells.


Products: Transfection Reagents

Summary of Reagents


Designed as a siRNA pDNA transfection reagent it can be used for either siRNA knockdown, plasmid DNA transformation or siRNA pDNA co-delivery. It is particularly suitable for mesenchymal stem cells from cord blood and bone marrow, and highly differentiated cells such as smooth muscle cells, and endothelial cells.


Download Information: All-Fect Brochure

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Benefits of the reagent:

  • High transfection efficiency

    Provides 2 to 3-fold higher efficacy in the presences of serum

  • Simple Protocol

    No need to change tissue culture medium during transfection

  • Toxicity

    Less toxic compared to commercial transfection reagents, leading to better retention of normal cellular physiology


  • Hsu and Uludağ. Biomaterials (2012) 33: 7834-7848.
  • Remant Bahadur et al., J. Materials Chemistry B (2015) 3: 3972-3982.
  • Wang et al., J. Surgical Research (2013) 183: 8-17.
All-fect transfection of cells GFP

Images: Typical performance of All-Fect for transfecting cord-blood derived mesenchymal stem cells with a GFP plasmid. The study compared the performance of All-Fect against a leading lipofection reagent under conditions optimized for each reagent. The extent of transgene expression was quantitated by flow cytometry, based on the extent of EGFP expression in arbitrary units. Typical fluorescent micrographs showing GFP expression after transfection are provided on the top.

Feature of Products

We developed broadly acting transfection reagents to modify mammalian cells with plasmid DNA, siRNA, mRNA and other nucleic acids. The foundation of our transfection reagents is based on cationic lipopolymers with optimal balance of cationic charge and hydrophobic (lipid) group. By systematically altering the polymeric backbone and the nature of lipid group, a library of transfection reagents has been generated. Some of the transfection reagents are broadly acting, functioning in different cell types with different nucleic acids. Others display high specificity, where they are exceptionally effective in particular cell types for particular nucleic acid delivery.

The key advantages of our delivery vehicles are:

  • Multivalent interactions with nucleic acids leading to strong binding of cargo that withstand disruptive forces in transit through cell membranes.
  • Synergistic effects due to cationic and lipidic binding that coat the cargo and protect it from nucleases.
  • Lipidic moieties that enhance interactions with cell memblane and internalization.
  • pH buffering capacitythat facilitate escape of cargo from endosomes.
  • Tailored formulations to free nucleic acids once internalized in cytoplasm.

The transfection reagents have been optimized for the following cell models and applications:

Primary Cells

Attachment Dependent

  • VSMC (Vascular Smooth Muscle Cells)
  • HUVEC (Human Umbilical Vein Endothelial Cells)
  • Human Foreskin Fibroblast Cells
  • BMSC (Bone Marrow Stromal Cells)
  • Human Myoblasts
  • Rat Primary Sympathetic Neurons

Attachment Independent

  • UCB-MSC (Umbilical Cord Blood Derived Mesenchymal Stem Cells)
  • BM-MSC (Bone Marrow Derived Mesenchymal Stem Cells)
  • Mononuclear Cells from Leukemia Patients
  • Mononuclear Cells from Normal Human Blood
Cell Lines

Attachment Dependent

  • 293-T (Kidney Fibroblast Cells)
  • MDA-231 (Breast Cancer Cells)
  • MDA-436 (Breast Cancer/ Melanoma Cells)
  • MDA-468, Sum-149PT, MCF-7 (Breast Cancer Cells)
  • A549 (Human Lung Cancer Cells)
  • MDCK (Kidney Epithelial Cells)
  • HCT-116 (Human Colon Cancer Cells)
  • C2C12 Myoblast Cells
  • MC3T3-1 (Preosteoblast Cells)
  • Green Monkey Vero Cells

Attachment Independent

  • U-937 (Human Lymphoma Cells)
  • K562 (Chronic Myeloid Leukemia Cells)
  • KG1, KG1A and THP-1 (Acute Myeloid Leukemia Cells)
  • Jurkat T-Cells
Animal Models
  • Systemic and local injection of RNAi mediator siRNA
  • Local injection of pDNA and mRNA expression vectors

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Background of Service

Our R&D Focus

One area of focus for RJH Biosciences is to implement RNA interference (RNAi) via delivery of short interfering RNA (siRNA). Our initial therapeutic application is blood cancers, while recognizing that the RNAi activity can be implemented in the treatment of a large range of human cancers and other diseases. Another focus is direct administration of plasmid DNA (pDNA) to express therapeutic proteins in situ, with applications in immunotherapy.

Why study blood cancers and immunotherapies with nucleic acid therapeutics?

There are three types of blood cancers: Leukemia, lymphoma, and myeloma. Leukemia is characterized by highly proliferating, abnormal white blood cells [1]. Lymphoma and myeloma are respectively cancers of the lymphatic system and plasma cells which greatly effect the immune system [2,3]. These three cancers are difficult to treat and the current treatments are limited in efficacy, especially at the end stage of the disease.

The use of nucleic acid-based therapeutics can eradicate these cancers in two primary ways, with RNAi technology and cell-based immunotherapy. The use of RNAi is being increasingly explored in the treatment of the blood cancers. Polynucleotides such as siRNA has aided the downregulation of oncogenes and can be designed to support specific abnormalities in individual patients, making it a ‘personal’ strategy with a universal technological design [4]. Due to siRNA’s potential in blood cancer therapies, we are currently focusing on siRNA therapeutics in our R&D projects, targeting disease-driving oncogenes and inducing apoptosis in the malignant cells.

Another strategy that is being explored for treating blood cancers is the use of immunotherapy. Immunotherapeutic strategies include the use of antibodies, stem cell transplants, cytokines, small molecules among others [5]. However, a more recent approach is genetic therapy by using engineered cells, also known as Cell Transfer Therapy. This method works by taking patients’ own immune cells such as but not limited to T-cells, B-cells, and NK cells. The immune cells genome is engineered to support various therapeutic strategies that may involve neoantigen expression and presentation on immune cell surface, and then are reintroduced into the host [5]. The modified cells are ultimately designed to target and remove the malignant cells. This strategy is highly advantageous as T-cells can ‘seek’ and destroy the malignant cells in the blood system. While this approach has been promising in blood cancers, it can be also used in other solid cancers. As the foundation of immunotherapy relies on nucleic acid introduction into patient cells, efficient delivery of nucleic acids is imperative for success. Our transfection reagents offer the best in class vehicles to undertake such a delivery.

Nanomedicine based on nucleic acid therapeutics is a large component to personalized cancer therapies and immunotherapies. The RJH Biosciences strives to provide quality transfection reagents, whether it involves the delivery of our own nucleic acid candidates or our customers’.

Jean, C. and Dick, J. (2005) Cancer stem cells: lessons from leukemia. Trends in cell biology. 15, 494-501.
Woods, N. et al. (2006) Therapueti gene causing lymphoma. Nature. 440, 1123.
Mahindra, A. et al. (2012) Latest advances and current challenges in the treatment of multiple myeloma. Nature Reviews Clinical Oncology. 9, 135-143.
Uludağ, H. et al. (2016) Current attempts to implement siRNA-based RNAi in leukemia models. Drug Discovery Today. 21, 1412-1420.
Zou, W. (2006) Regulatory T cells, tumour immunity and immunotherapy. Nature Reviews Immunology. 6, 295-307.

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Product List

Product List

Product Name Description Size
All-Fect pDNA, siRNA and co-delivery reagents for a broad range of cells 0.75 ml/1.5 ml
Prime-Fect Reagent of choice for tough to transfect primary and stem cells 0.75 ml/1.5 ml
Leu-Fect A & B Specialized reagents leukemia and suspension cells 0.75 ml/1.5 ml
Trans-Booster Designed to enhance the transfection efficiency of DNA/mRNA in attachment dependent and suspension-growing cells 0.75 ml/1.5 ml
In Vivo DNA-Fect Effective for pDNA delivery in animal models and cells 0.75 ml/1.5 ml
In Vivo RNA-Fect Effective systemic delivery of siRNA in animal models 0.75 ml/1.5 ml
mRNA-Fect Highly effective transfection reagent optimized for mRNA delivery 0.75 ml/1.5 ml
CRISP-Fect Highly effective transfection reagent optimized for ribonucleoprotein (RNP) delivery to both attachment-dependent and suspension-growing cells. 0.75 ml/1.5 ml

Transfection Reagent Selection Guide

The table below summarizes optimal transfection reagents for nucleic acids in different cell types.
The efficiency of the transfection reagents was assessed by using plasmid DNA (pDNA), short interfering RNA (siRNA), and messenger RNA (mRNA).
Where the transfection reagent was suitable for both pDNA or siRNA delivery, it was indicated with pDNA/siRNA.

Cell Type All-Fect Trans-Booster Leu-Fect-A Leu-Fect-B Prime-Fect mRNA-Fect CRISP-Fect
Primary Cells
Umbilical Cord Blood Derived Mesenchymal Stem Cells (UCB-MSC) pDNA pDNA
Bone Marrow Derived Mesenchymal Stem Cells (BM-MSC) pDNA pDNA
Vascular smooth muscle Cells (VSMCs) pDNA
Human Umbilical Vein Endothelial Cells (HUVECs) pDNA
Mononuclear Cells from CML patients (MNC) pDNA
Human Foreskin Fibroblast Cells pDNA
Rat Primary Sympathetic Neurons pDNA mRNA
Cell Lines
Kidney Fibroblast Cells (293-T) pDNA pDNA
Breast Cancer Cells (MDA-MB-231) pDNA
Kidney Epithelial Cells (MDCK) siRNA
Breast Cancer/Melanoma Cells (MDA-MB-436) pDNA
Breast Cancer Cells (MDA-MB-468) siRNA
Breast Cancer Cells (Sum-149PT) pDNA
Breast Cancer Cells (MCF-7) pDNA pDNA
Human Lymphoma Cells (U-937) pDNA pDNA
Chronic Myeloid Leukaemia Cells (K562) siRNA pDNA
Acute Myeloid Leukemia Cells (KG1 and KG1A) siRNA siRNA
Acute Myeloid Leukemia Cells (THP1) siRNA siRNA mRNA
Human Lung Cancer Cells (A549) siRNA
Human Colon Cancer (HCT-116) siRNA siRNA
Human Myoblasts ASO
Jurkat Cells pDNA mRNA RNP

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Example of Use

Feedback from independent researchers

Implementing CRISPR-Cas9 Technology using Transfection Reagents from RJH Biosciences
Killing Lung Cancer A549 Cells with RJH Reagents and Cytotoxic siRNAs
mRNA Transfection of Endothelial Cells with RJH Reagents
Transfecting Human Astrocytes with RJH Reagents: GFP Expression and siRNA Uptake
Transfection of A549 Lung Cancer Cells with RJH Reagents to Silence Tumor Suppressor p53 Expression
Transfecting Colon Cancer HCT-116 Cells with RJH Reagents to silence Polynucleotide Kinase 3′-Phosphatase (PNKP) Expression
Use of RJH Transfection Reagents in Antisense Oligonucleotide Delivery
Reagents for pDNA and siRNA delivery for TNBC cells

Application Notes

Reagents for Antisense Oligonucleotide (ASO) Delivery
Comparing Lipofection to RJH Reagents
mRNA Modification of PBMCs
mRNA-Fect Transfection Reagent to Deliver mRNA in Breast Cancer Cells
Transfecting Triple-Negative Breast Cancer MDA-MB-231 Cells with Plasmid DNA and siRNA by using ALL-Fect and Prime-Fect
Use of RJH Transfection Reagents in Co-Delivery of Plasmid DNA and short interfering RNA
RJH Transfection Reagents in siRNA Library Screens
microRNA Delivery to Leukemic Cells
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As the first step, we suggest consulting the ‘Transfection Reagent Selection Guide’ to find out if the cells of interest have been previously tested. If your cell type is not on the list, we suggest using the reagents optimized for similar cells (i.e., attachment-dependent/suspension or established cell line/primary cells), or contact us by e-mail for an informed suggestion.

We recommend small scale preliminary experiments to optimize the performance of our transfection reagents. As with all transfection reagents, the complex formation, cell seeding density, and culture and incubation conditions will affect the final performance. Please consult our technical sheet on optimizing transfection for this purpose (link).

Our transfection reagents are compatible with a wide range of serum-free media, including DMEM, RPMI, MEM and others. Complexes are expected to be functional in serum-containing medium.

The recommended nucleic acid to transfection reagent ratio ranges from (w/w) 1:1 for relatively toxic reagents to 5-20:1 for biocompatible reagents. This ratio should be optimized for each application. For suggested ranges for each reagent, please consult the specific reagent manual.

No. It not necessary to remove the complexes from treated cells. Complexes can be left in culture with cells until end-point analysis.

Depending on the application, incubation times may vary from 2 hours to 24 hours. It may be possible to centrifuge the treated cells to accelerate the transfection process and minimize the complex incubation times.

Transfection efficiency can be determined by different approaches. Reporter genes, such as GFP or RFP, are convenient ways to assess transfection by using microscopy or flow cytometry techniques. Direct assessment of the induced gene product (e.g., by ELISA, western blot), or silenced gene expression (e.g., protein levels or mRNA levels by PCR) are important. One can also use functional outcomes as an indirect measure of transfection, although care must be paid to complicating factors in this case. In all studies, we recommend employing a control (i.e., non-active) agent similar in nature to the nucleic acid being investigated.

A methodical analysis of the factors contributing to transfection, as outlined in the ‘Technical Tips to Improve Transfection’, is a good place to start. Other resources to improve transfection efficiencies can be found in our ‘suggested reading’ tab. We are here to help as well, so do send us a message/e-mail to see how we can assist you.

All transfection reagents display a certain extent of cytotoxicity on cells, depending on the amount used. The key is to achieve transfection without disrupting the physiology of the cells significantly. One can minimize exposure time to transfection complexes, speed up the transfection process by centrifugation and optimize reagent/nucleic acid concentrations to eliminate unnecessary exposure. The purity of the nucleic acid is also important to eliminate unforeseen toxicities.

These are important criteria that affect the transfection efficiency. In general, transfection efficiency decreases with increased cell density and passage number, as cells settle in senescence. For other factors affecting transfection efficiency, please consult ‘Technical Tips to Improve Transfection’.

The recommend storage temperature is at 4 degC (short term) or -20 degC (long term). The reagents are designed to be stable for 1 year under these conditions.

We recommend to use 1:5 ratio of nucleic acid to transfection reagent, so that 1 mL vial (1 mg) is suitable for 200 transfections of 1 mg nucleic acid. However, optimal ratio may change depending on the application and cell type.

Yes. Our in vivo transfection reagents display broad activities so that they could be effective under in vitro conditions.

It is likely for our transfection reagents to work with different nucleic acids. This is not universally applicable, but most reagents seem to handle different nucleic acids. ALL-Fect transfection reagent can handle both DNA and RNA.

Our reagents are based on an optimal balance of cationic charge and hydrophobicity. They are polymeric in nature that interact with nucleic acids via multivalent interactions. These reagents provide effective condensation of anionic charge of nucleic acids, while displaying little toxicity on mammalian cells.

To indicate the source of the transfection reagent, you can state the reagent name and that it was obtained from RJH Biosciences Inc. (Edmonton, AB, Canada).

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Please contact us for any inquiries, questions, or information requests.
Tokyo Future Style, Inc.
TEL:029-851-9222 FAX:029-851-9220