Transfection Reagents and Plasmid DNA
Transient transfection is a method of choice to modify the expression of specific gene(s) in eukaryotic cells. It requires three main components to create a transfection mix which are the nucleic acids (pDNA, mRNA, siRNA, etc), the transfection reagents and a complexation buffers/media.
The delivery system needs to be carefully selected based on the application and its challenges:
- Gene and Cell Therapy rely on the use of modified viruses as vectors to transfer genes into cells that they are able to infect. Production of viral vectors is dependent on efficient transfection of one to several plasmid DNA containing the necessary viral genome to produce recombinant viral particles.
- RNA/DNA Therapeutics using lipid nanoparticles (LNPs), polymers or even liposomes as a delivery system to target different organs via different administration routes (in vivo) depending on the therapeutic purpose.
- Protein Production (antibodies, peptides or proteins) that requires a screening phase using transient transfection of one or several plasmid DNA in CHO or HEK cell lines before moving to stable expression system at larger scale.
- Life Science Research that requires specific transfection system depending on various type of nucleic acids, cells, media and support of cell culture.
Choose among a broad range of powerful transfection reagents designed for specific application to be successful in your projects. Transfection reagents are ready-to-use with a pre-optimized protocol to ensure high transfection efficiency while preserving cell viability and morphology.
We assist academics, biotech and Pharma companies from R&D to manufacturing with end-to-end nucleic acid services, covering plasmid design, plasmid engineering, plasmid manufacturing (research to GMP), plasmid sequencing and process development and manufacturing of new biologics (viral vectors, recombinant proteins, antibody fragments, biosimilars and biobetters).
Transfection Reagents and Plasmids
Explore Our Transfection Reagents and Plasmids
Take a look at our innovation solution that offers gold-standard products and services from research to commercialization that enhance the quality and reliability of:
Viral Vector for CGT
The development of robust, scalable, cost-effective processes that yield high titers of high-quality viral vectors is crucial to the successful commercialization of safe and efficacious gene therapies that require large and/or multiple doses to treat more prevalent diseases. The successful commercialization of viral vector-based therapies relies on GMP-compliant and scalable, industrialized production platforms to rapidly advance high-quality products to market while also ensuring patient safety.
The most utilized approach to AAV (adeno-associated virus) and lentivirus (LV) production is through the transient transfection of mammalian cell lines, such as HEK293 or derivatives, followed by viral vector harvest, downstream purification processes and analytics for final product testing.
Since viral vector production is a significant cost driver in AAV and LV-based gene therapies, it is prudent to optimize each step of the upstream manufacturing process already at the process development phase to ensure compliance of critical raw materials (eg. plasmid DNA, transfection reagents), maximize yield and product quality while minimizing costs.
RNA|DNA Therapeutics
Non-viral in vivo delivery reagents are the most powerful alternative to viral vectors for nucleic acid delivery. They offer substantial advantages in terms of reliability, safety and costs for nucleic-acid based therapies. Different quality grades of in vivo transfection reagents are available for proof concept studies, preclinical studies and clinical trials.
Protein Production
Transient production of recombinant protein or monoclonal antibodies is an important step in research to study the behavior of a protein of interest in a cell, to screen and candidate proteins in early stages of therapeutic research and to enable modulation of molecular mechanism in many other fields of application.
Our goal is to help you make this protein production step as seamless as possible by optimizing the genetic engineering of your plasmid of interest and the transient expression system, whether bacterial, yeast or mammalian cell-based to meet your requirements for protein yield, function and quality.
What we can provide:
- Solutions for transient protein production in mammalian cells: we offer ready to use cost-effective transfection solutions (FectoPRO reagent) for transient protein production in mammalian cells, mainly HEK-293 and CHO cell lines, as well as plasmid engineering service to generate any plasmid encoding for your protein of interest.
- CDMO protein manufacturing service in bacteria and yeast: development of production methods for recombinant proteins and manufacturing of proteins and antibody fragments to fit your needs from 100 mg to 50g (Research, GLP and GMP grade).
Life Science Research
Explore our extensive range of life science delivery solutions for scientists, including ready-to-use transfection reagents suitable for most mammalian primary cells and cell lines, as well as off-the-shelf plasmids. These products ensure that you can conduct your research smoothly and rely on the following approaches:
- A range of best-in-class transfection reagents to study the function of a protein of interest, its role in a signaling pathway, perform genome editing using plasmid-based or RNA-based proven methods, and to conduct cost-effective RNA interference to transfect guide RNA and express Cas9 protein in mammalian cells.
- Tailored transfection solutions: Choose from our selection of specialized reagents, including jetPRIME®, INTERFERin®, and more, for targeted gene expression, genome editing, and RNA interference studies. Request samples and benefit from our scientific support to optimize your experiments and assist you during your evaluation phase.
pDNA Services
Our online plasmid design platform offers a unique intuitive interface to modularly create any kind of plasmid, to fit your experimental needs. This online portal is also the entry-point to our plasmid building platform operated with the e-Zyvec® DNA assembly technology. This allows us to robustly assemble the desired plasmids, regardless of sequence complexity. All our products can also be up-scaled from discovery grade, to R&D, to HQ and to GMP grades.
Our pDNA Services:
- Plasmid Engineering Service
- Whole Plasmid Sequencing Service
- Easy Plasmid Service
- Plasmid Manufacturing Service
Service Datasheets
Easy Plasmid Service - Plasmid Designed to Cater To Your Specific Requirements, Ensuring a Seamless And Cost-Effective Experience
PDF | 153.1 KBTailor-Made Plasmid Service - Engineering and Production Services for Plasmids From Discovery to GMP-Grade
PDF | 257.8 KBLooking for Guided Support
You can find certificate of analysis for all product range in Sartorius Portal. You can access them by creating an account.
From starting transfection, changing plasmid DNA construct, working with a new cell line or isolated primary cells can require fine-tuning. Our transfection experts are always available to help you.
Contact Support Team
The Who, The Why and How of Transfection?
Scientists rely on transfection as a powerful technique to modulate gene expression in eukaryotic cells in vitro and in vivo. Transfection can be used from researchers in universities or research institutes to engineers in Biotechnology or Pharmaceutical companies.
Transfection is the process that allows exogenous nucleic acids to bypass the cell membrane to enter into cells. Exogenous nucleic acids commonly used are plasmid DNA, RNA, siRNA and oligonucleotides. Once delivered into cells, nucleic acids modulate gene expression by driving overexpression or silencing of a gene of interest.
Gene overexpression is an indispensable tool for several applications, from understanding the role of gene of interest (gene studies, high-throughput screening), to the production of biologics such as antibodies (protein production) and recombinant viral particles, particularly for therapeutic purposes (virus production for gene & cell therapy).
Gene silencing is a method used to prevent expression of a gene of interest. The expression of a gene can be partially reduced (gene knockdown) or completely blocked (gene knockout). Because any gene can potentially be targeted, gene silencing is a prevalent technique used to develop gene-based therapies to address monogenic pathologies, cancer and in immunotherapy strategies.
Transfection of nucleic acids is used to transiently or stably modifed cells by overexpressing or silencing specific gene(s). Several methods can be used to performed transfection that are generally divided in two different categories: Chemical and physical. There are several physical methods that exist such as electroporation, sonoporation or microinjection but these processes are complex and relatively toxic for mammalian cells. To solve these issues, chemical-mediated transfection offers a great alternative: easiness of use, high transfection efficiency and excellent cell viability. Chemical transfection are typically performed using cationic polymers or lipids that will protect the anionic nucleic acids.
- Encapsulation of Genetic Material with Transfection Reagent
Nucleic acids are negatively charged due to their polyphosphate backbone and are thus able to interact with positively charged transfection reagents (polymers or lipids). This results in the formation of transfection complexes or nanoparticles, which protect nucleic acids from nuclease-mediated degradation. - Cellular Uptake of Nanoparticle
Most cells express negatively charged heparan sulfate proteoglycans on the external surface of their cell membrane, with which positively charged transfection complexes are able to interact. This interaction is key to trigger cellular uptake via an endocytosis process. - Release into the Cytosol and if Needed Transport into the Nucleus for Transcription
Upon cellular uptake, transfection complexes are sequestrated into intracellular vesicles. Our transfection reagents are able to induce the release of the nucleic acids into the cytoplasm through vesicle membrane rupture or fusion. Most nucleic acids (oligonucleotides, siRNA, mRNA, etc) stay in the cytoplasm where they are active. In case of gene transfer, plasmid DNA is transported into the nucleus for transient expression) which can become permanent after genome integration (stable expression).