Cell transfection is a technique used to introduce exogenous nucleic acids (such as DNA, mRNA, siRNA, miRNA, etc.) into cells, and it plays a critical role in modern biomedical research. With the deepening of scientific studies, the demand for different cell types has become increasingly diverse. From commonly used HEK293 cells to specialized tumor cells and primary cells, each cell type has unique characteristics and requirements. Similarly, different types of nucleic acids require specific transfection strategies to ensure efficient and safe gene delivery. Therefore, selecting the right transfection reagent is essential for experimental success.
Chemical transfection reagents are indispensable tools in cell biology research. They deliver nucleic acids such as DNA, mRNA, siRNA, and miRNA into cells through different mechanisms to achieve gene expression, silencing, or functional studies. Currently, the most commonly used chemical transfection reagents include liposomal transfection reagents, PEI-based reagents, and calcium phosphate reagents, each with its own advantages and application scenarios:
Table 1. Differences Between Common Chemical Transfection Reagents
|
Product Type |
Working Principle |
Advantages |
Disadvantages |
|
Liposomal Transfection Reagents |
1. Complex Formation: Liposomal reagents contain cationic lipids, whose positively charged head groups electrostatically bind to the negatively charged phosphate groups of nucleic acids, forming lipid–nucleic acid complexes. 2. Binding to Cell Membranes: Due to the positive charge of the complexes, they can adsorb onto the negatively charged cell membrane. 3. Endocytosis: Cells internalize the complexes through endocytosis, forming endosomes. 4. Endosomal Escape: Liposomes are often designed with a “proton sponge” effect, enabling cationic lipids to buffer the acidic endosomal environment, destabilizing the endosomal membrane and releasing the complexes into the cytoplasm. 5. Nucleic Acid Release: For siRNA, miRNA, or mRNA, functional activity occurs in the cytoplasm. For DNA, nuclear entry is required for expression. 6. Gene Expression: Once in the nucleus, DNA can be transcribed into mRNA and translated into protein, achieving exogenous gene expression. |
1. High transfection efficiency in various cell lines, including hard-to-transfect primary cells. 2. Broad applicability to both adherent and suspension cells, and to different nucleic acids (DNA, siRNA, miRNA, etc.). 3. Can be used in serum-containing medium without special serum removal. |
1. Cytotoxicity, especially at high concentrations or prolonged exposure. 2. Higher cost compared to traditional methods such as calcium phosphate transfection. 3. Limited in vivo application due to rapid serum clearance, lung accumulation, strong inflammatory response, and high toxicity. |
|
PEI Transfection Reagents |
1. Complex Formation: PEI is a highly cationic polymer that binds to negatively charged DNA via electrostatic interactions between amines on PEI and phosphate groups of DNA. 2. Membrane Attachment: PEI–DNA complexes adhere to the negatively charged cell membrane. 3. Endocytosis: Cells internalize the complexes via endocytosis, forming endosomes. 4. Endosomal Escape: PEI exhibits a “proton sponge” effect; protonation of amines in acidic endosomes draws in ions and water, causing swelling and rupture, releasing complexes into the cytoplasm. 5. DNA Release and Expression: Once released, PEI–DNA binding dissociates, allowing DNA to enter the nucleus for transcription and translation. |
1. High transfection efficiency due to stable complex formation. 2. Lower cytotoxicity compared to other cationic polymers. 3. Cost-effective, especially for large-scale viral vector production. 4. Available in GMP grade for clinical and commercial manufacturing. |
1. Less versatile—mainly suited for DNA delivery; limited application for mRNA, siRNA, or miRNA. 2. High efficiency mainly in HEK293 cells; less effective in hard-to-transfect cells. |
|
Calcium Phosphate Transfection Reagents |
1. Formation of Calcium Phosphate–DNA Co-precipitates: In phosphate-containing HEPES buffer, plasmid DNA binds to calcium ions to form co-precipitates via electrostatic interactions. 2. Cellular Uptake: The precipitates adsorb to the cell surface and are internalized via endocytosis. Particle size and quality directly affect efficiency. 3. DNA Release and Expression: Once inside, DNA is released and can be transiently expressed or stably integrated into the genome. |
1. Low cost, suitable for budget-limited labs. 2. Simple and easy to perform. 3. Applicable for both transient expression and stable cell line generation. |
1. Efficiency is highly variable and sensitive to factors such as pH and DNA purity. 2. Limited to certain cell types (mainly HEK293 for protein expression and viral packaging); not suitable for primary cells. |
With its strong R&D and manufacturing expertise, Yeasen Biotech continuously optimizes formulations for both DNA and RNA transfection reagents and improves production processes. The company offers a diversified product portfolio based on cationic lipids and cationic polymers, designed to meet the wide-ranging needs of research institutions and enterprises.
Advantages include:
Wide applicability: High-efficiency transfection of plasmid DNA, siRNA, miRNA, and mRNA
High transfection efficiency: Over 90% efficiency, suitable for multi-plasmid co-transfection
Multi-cell line validation: Verified in 40+ cell types with excellent results
Broad application scenarios: Stable cell line generation, transient protein expression, AAV and LV packaging
High citation frequency: 400+ high-impact publications, total IF exceeding 3000
GMP-grade products: Supporting commercial-scale production and regulatory submissions
How to Choose the Right Transfection Reagent
Given the unique requirements of different cell types and nucleic acids (DNA, mRNA, siRNA, etc.) for optimal transfection efficiency and conditions, choosing the right reagent is essential for success. The selected reagent should maximize efficiency while minimizing cytotoxicity to ensure data accuracy and reproducibility. Yeasen offers a series of optimized products for various applications, ensuring researchers can find the most suitable solution for their specific needs.
Product Information
|
Cat. No. |
Product Name |
Cell Type |
Nucleic Acid Type |
Application |
|
40802ES |
Common cells |
DNA |
High-performance liposomal reagent for efficient transfection in multiple cell types |
|
|
40804ES |
Common cells |
Lentivirus |
Enhances lentiviral infection |
|
|
40806ES |
Common cells (hard-to-transfect) |
siRNA, miRNA, ASO |
Dedicated siRNA/miRNA reagent with high knockdown efficiency |
|
|
40809ES |
Common cells (hard-to-transfect) |
mRNA |
High-efficiency mRNA delivery in multiple cell types |
|
|
40815ES |
Common cells |
DNA |
Broadly used for protein expression and viral packaging |
|
|
40816ES |
Common cells |
DNA |
General-purpose viral packaging transfection reagent |
|
|
40820ES |
Common cells |
DNA |
— |
|
|
40823ES |
Common cells |
DNA |
AAV-specific packaging transfection reagent |
|
|
40824ES |
Common cells |
DNA |
GMP-grade AAV-specific transfection reagent |
Product Performance
Single Plasmid Transfection
In a 6-well plate system, Yeasen 40802ES and competitor reagents were used to transfect a GFP plasmid into HEK293 cells. GFP expression was observed 48 h post-transfection under a fluorescence microscope.
Dual Plasmid Transfection
Cell type: HEK293;Format: 12-well plate, transient transfection;Total plasmid amount: 1 μg;Transfection reagent: 3 μL
Customer Case Studies
Bacterial protein induces autophagosome formation
Cells: Hela
Transfected plasmid: Bacterial effector protein A
LC3II: Autophagosome marker protein
Mitotic spindle structure
Cells: Hela
Transfected plasmid: Flag-tubulin
Nuclear staining: Hoechst33342
Multi-Cell Line Validation
|
Cell Type |
Cell Type |
Cell Type |
Cell Type |
Cell Type |
|
293FT |
Caco-2 |
HEK 293T |
LM3 |
NIH-3T3 |
|
293T |
CHO-K1 |
HEK293 |
MCF10A |
PC12 |
|
3T3 |
COS-7 |
HeLa |
MCF-7 |
Raw264.7 |
|
5-8F |
DF-1 |
Hep 3B |
MDA-MB-231 |
RKO |
|
A549 |
H520 |
Hepa1-6 |
MEF |
SGC-7901 |
|
BV-2 |
H9 |
HepG2 |
MKN-28 |
SMCC7721 |
|
C2C12 |
H9c2 |
HUVEC |
N2A |
Vero |
|
C6 |
HaCaT |
Lenti X-293T |
NCI-H1975 |
HCT116 |
|
WRL-68 |
THP-1 |
MDCK |
Hep2C |
More… |
High-Impact Literature Citations (Selected Examples)
1. Liang X, Gong M, Wang Z, et al. LncRNA TubAR complexes with TUBB4A and TUBA1A to promote microtubule assembly and maintain myelination. Cell Discov. 2024;10(1):54. Published 2024 May 21. doi:10.1038/s41421-024-00667-y. IF=33.5(40808ES)
2. Wang A, Chen C, Mei C, et al. Innate immune sensing of lysosomal dysfunction drives multiple lysosomal storage disorders. Nat Cell Biol. 2024;26(2):219-234. doi:10.1038/s41556-023-01339-x.IF=21.3(40802ES)
3. Liu H, Zhen C, Xie J, et al. TFAM is an autophagy receptor that limits inflammation by binding to cytoplasmic mitochondrial DNA. Nat Cell Biol. 2024;26(6):878-891. doi:10.1038/s41556-024-01419-6.IF=21.3(40802ES)
4. Wang WW, Ji SY, Zhang W, et al. Structure-based design of non-hypertrophic apelin receptor modulator. Cell. 2024;187(6):1460-1475.e20. doi:10.1016/j.cell.2024.02.004.IF=64.5(40802ES)
5. Ke J, Pan J, Lin H, et al. Targeting Rab7-Rilp Mediated Microlipophagy Alleviates Lipid Toxicity in Diabetic Cardiomyopathy. Adv Sci (Weinh). Published online June 5, 2024. doi:10.1002/advs.202401676.IF=15.1(40806ES)
6. Jiang L, Xie X, Su N, et al. Large Stokes shift fluorescent RNAs for dual-emission fluorescence and bioluminescence imaging in live cells. Nat Methods. 2023;20(10):1563-1572. doi:10.1038/s41592-023-01997-7.IF=48(40802)
7. Lou M, Huang D, Zhou Z, et al. DNA virus oncoprotein HPV18 E7 selectively antagonizes cGAS-STING-triggered innate immune activation. J Med Virol. 2023;95(1):e28310. doi:10.1002/jmv.28310.IF=20.69(40802ES)
8. Su J, Shen S, Hu Y, et al. SARS-CoV-2 ORF3a inhibits cGAS-STING-mediated autophagy flux and antiviral function. J Med Virol. 2023;95(1):e28175. doi:10.1002/jmv.28175.IF=20.69(40802ES)
9. Lu YY, Zhu CY, Ding YX, et al. Cepharanthine, a regulator of keap1-Nrf2, inhibits gastric cancer growth through oxidative stress and energy metabolism pathway. Cell Death Discov. 2023;9(1):450. Published 2023 Dec 12. doi:10.1038/s41420-023-01752-z.IF=7(40806ES)
10. Li X, Zhang Y, Xu L, et al. Ultrasensitive sensors reveal the spatiotemporal landscape of lactate metabolism in physiology and disease. Cell Metab. 2023;35(1):200-211.e9. doi:10.1016/j.cmet.2022.10.002.IF=31.373(40802ES)
11. Li X, Zhang Y, Xu L, et al. Ultrasensitive sensors reveal the spatiotemporal landscape of lactate metabolism in physiology and disease. Cell Metab. 2023;35(1):200-211.e9. doi:10.1016/j.cmet.2022.10.002.IF=31.373(40804ES)
12. Huang Y, Motta E, Nanvuma C, et al. Microglia/macrophage-derived human CCL18 promotes glioma progression via CCR8-ACP5 axis analyzed in humanized slice model. Cell Rep. 2022;39(2):110670. doi:10.1016/j.celrep.2022.110670.IF=8.8(40804ES)
13. Chai Q, Yu S, Zhong Y, et al. A bacterial phospholipid phosphatase inhibits host pyroptosis by hijacking ubiquitin. Science. 2022;378(6616):eabq0132. doi:10.1126/science.abq0132.IF=63.714(40802ES)
14. Liu R, Yang J, Yao J, et al. Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins. Nat Biotechnol. 2022;40(5):779-786. doi:10.1038/s41587-021-01112-1.IF=54.908(40802ES)
15. Chen S, Chen G, Xu F, et al. Treatment of allergic eosinophilic asthma through engineered IL-5-anchored chimeric antigen receptor T cells. Cell Discov. 2022;8(1):80. Published 2022 Aug 16. doi:10.1038/s41421-022-00433-y.IF=38.079(40804ES)