Transfection Reagents for DNA, RNA, siRNA, and CRISPR Delivery into Cultured Cells

Transfection reagents introduce DNA, mRNA, siRNA, and CRISPR/Cas9 components into cultured cells via chemical (cationic lipids/lipofection, PEI, calcium phosphate), physical (electroporation), or viral (transduction with polybrene/DEAE-dextran enhancement) methods. Reagent choice depends on nucleic acid type, cell type, and whether transient or stable expression is the goal — primary and stem cells often require empirical testing across chemistries. Electroporation targets 40–80% survival with 1–5 µg DNA per 10⁷ cells. MBP ships to labs across the United States, Canada, and internationally with PO and Quick Order at mbpinc.net.

Explore available transfection reagents or request a quotation by contacting customerservice@mbpinc.net. Our team can help identify the optimal delivery method for your cell type, nucleic acid cargo, and expression requirements.

Transfection Reagents

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What are transfection reagents?

Transfection reagents enable the delivery of exogenous nucleic acids — plasmid DNA, mRNA, siRNA/shRNA, and CRISPR/Cas9 ribonucleoprotein or plasmid components — into cultured cells for applications spanning gene expression studies, gene silencing, CRISPR-based gene editing, recombinant protein production, and stable cell line development. Delivery methods fall into three categories: chemical methods (cationic lipid/lipofection reagents, polymeric agents like polyethyleneimine/PEI, and calcium phosphate co-precipitation), physical methods (electroporation and biolistic particle delivery), and viral methods (transduction via lentiviral, adenoviral, or AAV vectors, the only category where "transduction" is the technically correct term). Each category includes multiple specific reagents with different optimal nucleic acid types, cell type compatibility, and transient-versus-stable expression outcomes.

What you will find:

DNAfectin™ Plus Transfection Reagent: innovative lipid-based reagent for rapid transfection in common adherent cell lines; fewer handling steps, and works in complete growth media
Susfectin™ Transfection Reagent: polymer-based reagents formulated for suspension cultures, providing highly efficient nucleic acid delivery
Transfection Reagent: designed for faster and more reliable concentration of lentiviral particles from cell culture supernatant

How to choose transfection reagents

Chemical: lipofection

Cationic lipid (lipofection) reagents form positively charged complexes with nucleic acids that interact with the negatively charged cell membrane, enabling entry primarily via endocytosis. Examples include DOTMA-based formulations and product lines such as Lipofectamine and X-tremeGENE, suitable for a diverse range of cell lines with DNA, small RNA, and CRISPR/Cas9 components. Lipid transfections adapt well to cost-effective and high-throughput formats but are generally cell-type-specific in optimal performance — a reagent validated on one line may need re-optimization for another.

Chemical: PEI and calcium phosphate

Polyethyleneimine (PEI) is a polymeric transfection agent that, like cationic lipids, forms complexes with DNA for cell membrane interaction — commonly used for high-yield transient transfection in suspension cultures for recombinant protein production. Calcium phosphate co-precipitation is a long-established method (Graham & van der Eb, 1973) that remains highly efficient for a wide range of cell types and molecules, suitable for both transient and stable transfection, though the process can be harsher on sensitive cell lines, leading to increased cell death in some cases.

Physical: electroporation

Electroporation exposes cells to electrical pulses that create temporary membrane pores, allowing direct entry of DNA, RNA, and other molecules — an easy, non-chemical technique yielding high efficiency across diverse cell types. Optimization centers on three parameters: pulse voltage, pulse width, and pulse number, with the goal of maintaining 40-80% cell survival. Electroporation requires roughly five-fold more cells and DNA than calcium phosphate methods, typically 1-5 µg DNA per 10⁷ cells, with good linear correlation between DNA amount and uptake.

Viral transduction enhancers

For viral transduction (lentiviral, adenoviral, AAV), polybrene and DEAE-dextran are polycations that minimize electrostatic repulsion between negatively charged viral particles and cell membranes, enhancing transduction efficiency. FBS has been shown to yield better transduction efficiency than bovine calf serum in some transduction protocols — a media-composition detail relevant to viral workflows specifically.

Selecting reagent for nucleic acid type

Some reagents are specifically optimized for plasmid DNA (Effectene, TransIT-X2), while others (Lipofectamine RNAiMAX) are designed for small oligonucleotides like siRNA. Co-transfection of multiple nucleic acid types may require protocols and ratios validated specifically for that combination rather than direct extrapolation from single-nucleic-acid optimization.

Pre-transfection cell preparation

Regardless of method, cells should be at least 90% viable and have recovered at least 24 hours post-subculture before transfection. Cell density must avoid both contact inhibition (too high, reducing uptake) and insufficient cell-cell contact (too low, reducing growth) — both extremes reduce efficiency.

Specifications context

Electroporation optimization targets 40-80% post-pulse cell survival across a parameter space of pulse voltage, width, and number, with typical DNA loading of 1-5 µg per 10⁷ cells showing linear uptake correlation within that range. Calcium phosphate transfection (Graham & van der Eb, 1973) remains a foundational method still in active use for both transient and stable transfection across many cell types. As of 2026, lipid-based reagents remain the default first choice for adherent cell line transfection due to ease of use, with electroporation and viral transduction reserved for hard-to-transfect primary/stem cells and applications requiring stable genomic integration respectively.

Contact the expert team at MBP and find premium transfection reagents for your lab today.

FAQ

Transfection reagents are chemicals and tools used to introduce exogenous nucleic acids (DNA, mRNA, siRNA, CRISPR/Cas9 components) into cultured cells. The three main delivery method categories are chemical (cationic lipids/lipofection, polymers like polyethyleneimine/PEI, calcium phosphate co-precipitation), physical (electroporation, biolistic particle delivery), and viral (transduction using lentiviral, adenoviral, or AAV vectors). Each method category includes multiple specific reagents and protocols suited to different cell types and experimental goals.

Lipofection relies on cationic lipid complexes that fuse with or are taken up by the cell membrane via endocytosis, delivering nucleic acids in a relatively gentle, chemically-mediated process — examples include DOTMA-based reagents and Lipofectamine/X-tremeGENE product lines. Electroporation uses electrical pulses to create temporary pores in the cell membrane, allowing direct entry of DNA, RNA, or other molecules — a non-chemical, physical method that can achieve high efficiency across many cell types but can be harsher, causing more cell death in sensitive lines.

Transfection refers to introducing nucleic acids into cells by non-viral means (chemical or physical methods). Transduction is the correct term specifically for virus-mediated gene delivery — using viral vectors (lentivirus, adenovirus, AAV) to carry genetic material into target cells. Viral vectors offer high transduction efficiency and can be tailored for specific cell types, with lentiviral vectors capable of integrating into the host genome for sustained expression, distinguishing transduction from the typically transient expression achieved by most chemical transfection methods.

Cell viability and health prior to transfection are critical — cells should be at least 90% viable and have recovered for at least 24 hours after subculturing before transfection. Cell density matters: too high causes contact inhibition and poor nucleic acid uptake, while too few cells may show poor growth without cell-to-cell contact. For lipid-mediated transfection, the DNA-to-reagent ratio and exposure time to the lipid-DNA complex affect both efficiency and toxicity. For viral transduction, actively dividing cells transduce more efficiently, and non-dividing cell types may require increased multiplicity of infection (MOI).

Yes — reagent choice often depends on the nucleic acid type and transfection complexity. Some reagents (such as Effectene and TransIT-X2) are specifically optimized for plasmid DNA transfection, while others (such as Lipofectamine RNAiMAX) are designed for transfecting small oligonucleotides like siRNA. Co-transfection (multiple nucleic acids simultaneously) may require reagents and ratios specifically validated for that use case rather than assuming a single-nucleic-acid protocol scales directly.

Electroporation is primarily governed by three electrical parameters: pulse voltage, pulse width, and pulse number. The optimization objective is to find a pulse combination that maintains 40-80% cell survival while achieving sufficient membrane permeabilization for nucleic acid entry. Electroporation requires roughly five-fold more cells and DNA compared to calcium phosphate-mediated transfection, with a typical guideline of 1-5 µg DNA per 10⁷ cells, and shows good linear correlation between DNA amount and uptake within that range.

Primary and stem cells are frequently challenging to transfect because they often have lower baseline transfection efficiency with standard reagents, may show greater sensitivity to reagent-associated toxicity, and can respond differently to different reagent chemistries — non-liposomal reagents have shown better efficiency in some primary human cells, while liposomal reagents like Lipofectamine have performed better in others, with no universal best choice. Empirical testing of multiple reagents on the specific primary/stem cell type is standard practice rather than relying on results from immortalized line validation data.

MBP supports purchase order and Quick Order procurement for transfection reagents — lipid-based reagents, polymers (PEI), electroporation-compatible buffers, and viral transduction enhancers (polybrene, DEAE-dextran) — for labs in the United States, Canada, and internationally. MBP is a registered vendor for Howard Hughes Medical Institute, Vanderbilt University, MD Anderson Cancer Center, and Eurofins. For sourcing specific reagent lines or volume pricing, contact MBP at mbpinc.net.

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