Cell transfection is a technique for introducing exogenous nucleic acids (such as DNA, mRNA, siRNA, miRNA, etc.) into cells, and it plays a crucial role in modern biomedical research. As scientific research continues to delve deeper, the demand for a variety of cell types has also become increasingly diverse, ranging from common HEK293 cells to special types of tumor cells and primary cells, each with its unique characteristics and requirements. Similarly, different types of nucleic acids also require specific transfection strategies to ensure efficient and safe gene delivery. Therefore, choosing the right transfection reagent is essential for the success of the experiment.
Chemical transfection reagents are indispensable tools in cell biology research, which introduce nucleic acids such as DNA, mRNA, siRNA, miRNA, etc., into cells through different mechanisms to achieve gene expression, silencing, or functional studies. Currently, commonly used chemical transfection reagents mainly include liposome transfection reagents, PEI transfection reagents, and calcium phosphate transfection reagents, each with its unique advantages and applicable scenarios.
Table 1: What are the differences among different chemical transfection reagents
|
Product Category |
Principle |
Advantages |
Disadvantages |
|
Liposome transfection reagent |
1. Liposome transfection reagents have positive-charged lipids that bind to negatively-charged nucleic acids to form lipid-nucleic acid complexes. 2. These complexes are attracted to the negatively-charged cell membrane. 3. The cell engulfs the complexes into endosomes through endocytosis. 4. The "proton sponge" effect of the lipids in the complexes prevents endosome acidification, leading to membrane destabilization and rupture, releasing the complexes into the cytoplasm. 5. Small nucleic acids like siRNA, miRNA, or mRNA can work in the cytoplasm, while DNA needs to enter the nucleus to be expressed. 6. Once in the nucleus, DNA is transcribed into mRNA and translated into proteins, expressing the foreign gene. |
1. Liposome transfection reagents offer high efficiency with various cell lines, including hard-to-transfect primary cells. 2. They are versatile, compatible with both adherent and suspension cells, and various nucleic acids like DNA, siRNA, and miRNA. 3. These reagents work well in serum-containing media, avoiding the need to remove serum and thus minimizing cell damage. |
1. Liposome transfection reagents can be toxic to cells, especially at high concentrations or over long periods, potentially affecting cell survival and function. 2. They are more expensive compared to traditional methods like calcium phosphate transfection. 3. Their in vivo application is limited due to serum clearance, accumulation in lung tissue, and potential to cause strong inflammatory responses and high toxicity. |
|
PEI transfection reagent |
1. PEI, a positively charged polymer, binds to negatively charged DNA to form PEI-DNA complexes through electrostatic interactions. 2. These complexes attach to the negatively charged cell membrane. 3. Cells engulf the PEI-DNA complexes through endocytosis, forming endosomes. 4. The "proton sponge" effect of PEI causes endosomes to swell and burst in the acidic environment, releasing the complexes into the cytoplasm. 5. In the cytoplasm, PEI-DNA bonds break, allowing DNA to enter the nucleus, where it is transcribed and translated into proteins. |
1. PEI transfection reagents form stable complexes with DNA, enhancing transfection efficiency. 2. They have lower cytotoxicity compared to other cationic polymers, preserving cell viability and function. 3. PEI reagents offer a cost-effective solution, especially for large-scale transfections like viral vector production. 4. PEI reagents are available in GMP-grade for clinical and commercial production, ensuring quality and compliance. |
1. PEI transfection reagents have limited versatility, primarily used for DNA and less effective with other nucleic acids like mRNA, siRNA, and miRNA. 2. They show high efficiency in specific cell types, such as HEK293 cells, for virus packaging and protein expression, but perform poorly with difficult-to-transfect cells. |
|
Calcium Phosphate transfection reagent |
1. DNA/calcium phosphate co-precipitates form in HEPES buffer with phosphate, where DNA's negative charge binds to positive calcium ions. 2. Cells engulf these co-precipitates, which stick to their surface, through endocytosis, a critical step for transfection efficiency. 3. Inside cells, the co-precipitates release DNA, which can lead to transient expression or stable integration into the genome. |
1. Calcium phosphate transfection is cost-effective, suitable for labs with limited budgets. 2. It's easy to perform, requiring simple steps and minimal technical skill. 3. It can be used for both transient protein expression and creating stable cell lines. |
1. Calcium phosphate transfection efficiency is variable and sensitive to factors like pH and DNA purity, which can lead to failed transfections, especially with impure DNA. 2. It's not suitable for all cell types, mainly used for HEK293 cells, and less effective for primary cells and some other cell types. |
Yeasen Biotech, leveraging its strong R&D capabilities and production technology team, continuously optimizes the formulations of DNA and RNA transfection reagents and improves production processes. The company has launched a diversified product line based on cationic liposomes and cationic polymers to meet the broad needs of research institutions and enterprises in the field of transfection reagents. These products cover various applications of transfection reagents and have the following advantages:
Broad Applicability: Capable of efficiently transfecting plasmid DNA, siRNA, miRNA, and mRNA.
High Transfection Efficiency: Cell transfection efficiency exceeds 90%, meeting the needs for co-transfection of multiple plasmids.
Verified Across Multiple Cell Lines: Demonstrated good transfection efficiency in over 40 different cell lines.
Wide Range of Applications: Used for stable cell line construction, transient protein expression, and AAV and LV viral packaging.
High Citation Frequency in Literature: Cited in over 400 high-impact publications, with a total impact factor exceeding 3000+.
How to choose a transfection reagent that suits your needs
Given the special requirements for transfection efficiency and conditions in different cell types and nucleic acids (such as DNA, mRNA, siRNA, etc.), selecting the right transfection reagent is crucial for the success of the experiment. Based on the specific needs of the experiment, choose a transfection reagent that can optimize transfection efficiency and minimize cytotoxicity to ensure the accuracy of experimental data and the robustness of experimental results. YEASEN Biotech offers a range of products optimized for different application scenarios, ensuring that you can find the most suitable transfection reagent for your research purposes and meet your specific experimental needs.
Case Presentation
In a 6-well plate system, GFP-expressing plasmids were transfected into HEK293 cells using YEASEN 40802ES transfection reagent and a competitor's transfection reagent. The GFP expression in transfected cells was observed under a microscope 48 hours post-transfection. The performance of YEASEN's transfection reagent was superior.
In a 12-well plate, HEK293 cells were transfected with a total of 1 μg of plasmid DNA; using 3 μL of transfection reagent, it was possible to successfully co-transfect two plasmids.
Customer Case
Validated across multiple cell lines, with a broader range of applications.
|
Cells |
Cells |
Cells |
Cells |
Cells |
|
293F |
Caco2 |
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… |
Product List
|
Product Name |
Product Specifications |
Product number |
|
Hieff Trans® Liposomal Transfection Reagent |
0.5 mL/1 mL/5×1 mL |
40802ES02/ES03/ES08 |
|
Hieff Trans®Calcium Phosphate Cell Transfection Kit |
200T |
40803ES70 |
|
Hieff Trans®Polybrene(hexadimethrine bromide) (10 mg/mL) |
500 μL/5×500 μL |
40804ES76/ES86 |
|
Hieff Trans® in vitro siRNA/miRNA Transfection Reagent |
0.5 mL/1 mL |
40806ES02/ES03 |
|
Hieff Trans® Universal Transfection Reagent |
0.5 mL/1 mL/5×1 mL |
40808ES02/ES03/ES08 |
|
Hieff Trans® mRNA Transfection Reagent |
0.1 mL/1 mL |
40809ES01/ES03 |
|
Hieff Trans®Polyethylenimine Linear(PEI) MW25000 |
1 g/5 g |
40815ES03/ES08 |
|
Hieff Trans®Polyethylenimine Linear (PEI) MW40000（rapid lysis） |
100 mg/1 g |
40816ES02/ES03 |
|
Hieff Trans®293 Transfection Reagent |
1 mL/10 mL/100 mL |
40818ES03/ES10/ES60 |
Cited in multiple high-impact publications, ensuring quality (partial list of cited references)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)
- 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)