Transfectamine™ mRNA Transfection Reagent

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
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Shipping	Standard overnight for United States, inquire for international

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Water

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Transfectamine™ mRNA Transfection Reagent is a powerful and versatile transfection reagent designed to introduce a higher amount of mRNA into eukaryotic cells, or more specifically, into animal cells. It delivers high transfection efficiency in a wide variety of adherent and suspension cell lines, including difficult-to-transfect cells. Nuclear uptake is not required, which results in faster protein expression than DNA transfection without the risk of genomic integration. The low toxicity of Transfectamine™ mRNA Transfection Reagent allows higher viability of transfected cells. Transfectamine™ mRNA Transfection Reagent does not require special medium and is easier to use compared to most of the commercial transfection reagents.

Example protocol

AT A GLANCE

Protocol Summary

Prepare cells for transfection.
Prepare the Transfectamine™ mRNA Transfection Reagent-RNA mixture.
Add the Transfectamine™ mRNA Transfection Reagent-RNA mixture to the cell culture.
Culture cells overnight.
Analyze transfection efficiency with an appropriate method.

CELL PREPARATION

Culture cells to ~ 90% confluency at the time of transfection.
Replace with fresh growth medium before transfection. For example, replace with 2 mL of medium per well for 6-well plates and 6 mL of medium for 10 cm plates.

PREPARATION OF WORKING SOLUTION

Transfectamine™ mRNA Transfection Reagent-RNA mixture

Mix 2.5 µg of mRNA with 200 µL of serum-free medium.
Add 7.5 µL of Transfectamine™ mRNA Transfection Reagent to Step 1.
Mix well and incubate at RT for 20 minutes.

Note: The ratio of Transfectamine™ mRNA Transfection Reagent to mRNA needs to be optimized for different cell lines. In general, Transfectamine™ mRNA Transfection Reagent (µL) to mRNA (µg) Ratio = (3 to 5 µL) to 1 µg.

Table 1. Sample protocol detail for 6-well plates as shown in the table below.

Component	6-well plate (per well)
Fresh culture medium	2 mL
Purified mRNA	~2.5 µg
Serum-free medium	200 µL
Transfectamine™ mRNA Transfection Reagent	~7.5 µL

SAMPLE EXPERIMENTAL PROTOCOL

Transfection Protocol

Add the Transfectamine™ mRNA Transfection Reagent -mRNA mixture to the culture plate and culture overnight.

Note: Recombinant protein expression can be detected as early as 8 hours after the transfection. Maximal expression level may be observed ~24 hours after the transfection.

Images

Figure 1. Transfection efficiency comparison (Upper panel) and cellular toxicity comparison (Bottom panel) in HeLa cells. HeLa cells were cultured in a 6-well plate to ~90% confluency. 2.5 µg of mRNA was transfected with Lipofectamine MessengerMAX and Transfectamine™ mRNA Transfection Reagent, respectively. Images were taken 18 hours after the transfection using a fluorescent microscope with the FITC channel (Upper panel). Although transfection efficiency was similar for Lipofectamine MessengerMAX and Transfectamine™ mRNA Transfection Reagent, most Lipofectamine MessengerMAX transfected samples were scrambled, whereas cells transfected with Transfectamine™ mRNA Transfection Reagent looked much healthier (bottom panel).

Figure 2. Cell viability comparison in HeLa cells. HeLa cells were incubated with Transfectamine™ mRNA Transfection Reagent and Lipofectamine MessagerMax, respectively, according to suggested protocol without mRNA. After 24 hours, cell viability in both groups was measured with Cell Meter™ Colorimetric WST-8 Cell Quantification Kit (Cat#22770). The higher absorbance at 460 represents more numbers of viable cells.

References

View all 50 references: Citation Explorer

Preclinical evaluation of CD8+ anti-BCMA mRNA CAR T cells for treatment of multiple myeloma.
Authors: Lin, Liang and Cho, Shih-Feng and Xing, Lijie and Wen, Kenneth and Li, Yuyin and Yu, Tengteng and Hsieh, Phillip A and Chen, Hailin and Kurtoglu, Metin and Zhang, Yi and Andrew Stewart, C and Munshi, Nikhil and Anderson, Kenneth C and Tai, Yu-Tzu
Journal: Leukemia (2021): 752-763

Sustained release of PKR inhibitor C16 from mesoporous silica nanoparticles significantly enhances mRNA translation and anti-tumor vaccination.
Authors: Zhang, Wei and Liu, Yi and Min Chin, Jas and Phua, Kyle K L
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2021): 179-187

A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair generating improved genetically stable human induced pluripotent stem cells.
Authors: Alvarez-Palomo, Ana Belén and Requena-Osete, Jordi and Delgado-Morales, Raul and Moreno-Manzano, Victoria and Grau-Bove, Carme and Tejera, Agueda M and Otero, Manel Juan and Barrot, Carme and Santos-Barriopedro, Irene and Vaquero, Alejandro and Mezquita-Pla, Jovita and Moran, Sebastian and Naya, Carlos Hobeich and Garcia-Martínez, Iris and Pérez, Francisco Vidal and Blasco, María A and Esteller, Manel and Edel, Michael J
Journal: Stem cells (Dayton, Ohio) (2021)

CD40 signaling augments IL-10 expression and the tolerogenicity of IL-10-induced regulatory dendritic cells.
Authors: Dawicki, Wojciech and Huang, Hui and Ma, Yanna and Town, Jennifer and Zhang, Xiaobei and Rudulier, Chris D and Gordon, John R
Journal: PloS one (2021): e0248290

mRNA Transfection of T-Lymphocytes by Electroporation.
Authors: Schwarze, Lea-Isabell and Fehse, Boris
Journal: Methods in molecular biology (Clifton, N.J.) (2021): 217-226

A Systematic Study of Unsaturation in Lipid Nanoparticles Leads to Improved mRNA Transfection In Vivo.
Authors: Lee, Sang M and Cheng, Qiang and Yu, Xueliang and Liu, Shuai and Johnson, Lindsay T and Siegwart, Daniel J
Journal: Angewandte Chemie (International ed. in English) (2021): 5848-5853

Expediting in vitro characterization of mRNA-based gene therapies via high-content fluorescent imaging.
Authors: Vigil, Toriana N and Zhang-Hulsey, Diana and Santos, Jose Luis and Patrick Hussmann, G
Journal: Analytical biochemistry (2021): 114259

Live-cell Imaging of Single-Cell Arrays (LISCA) - a Versatile Technique to Quantify Cellular Kinetics.
Authors: Reiser, Anita and Woschée, Daniel and Kempe, Simon Maximilian and Rädler, Joachim Oskar
Journal: Journal of visualized experiments : JoVE (2021)

Location of a single histidine within peptide carriers increases mRNA delivery.
Authors: He, Jiaxi and Xu, Songhui and Leng, Qixin and Mixson, A James
Journal: The journal of gene medicine (2021): e3295

PEGylation of poly(amine-co-ester) polyplexes for tunable gene delivery.
Authors: Grun, Molly K and Suberi, Alexandra and Shin, Kwangsoo and Lee, Teresa and Gomerdinger, Victoria and Moscato, Zoe M and Piotrowski-Daspit, Alexandra S and Saltzman, W Mark
Journal: Biomaterials (2021): 120780