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Transfectamine™ mRNA Transfection Reagent

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

  1. Prepare cells for transfection.

  2. Prepare the Transfectamine™ mRNA Transfection Reagent-RNA mixture.

  3. Add the Transfectamine™ mRNA Transfection Reagent-RNA mixture to the cell culture.

  4. Culture cells overnight.

  5. Analyze transfection efficiency with an appropriate method.

CELL PREPARATION

  1. Culture cells to ~ 90% confluency at the time of transfection.

  2. 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

  1. Mix 2.5 µg of mRNA with 200 µL of serum-free medium.

  2. Add 7.5 µL of Transfectamine™ mRNA Transfection Reagent to Step 1.

  3. 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.

Component6-well plate (per well)
Fresh culture medium2 mL
Purified mRNA~2.5 µg
Serum-free medium200 µL
Transfectamine™ mRNA Transfection Reagent~7.5 µL

SAMPLE EXPERIMENTAL PROTOCOL

Transfection Protocol

  1. 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.

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
YAF2-mediated YY1-Sirtuin6 interactions responsible formitochondrial down-regulation in aging tunicates.
Authors: Kawamura, Kaz and Higuchi, Takuma and Fujiwara, Shigeki
Journal: Molecular and cellular biology (2021)
LipoParticles: Lipid-Coated PLA Nanoparticles Enhanced In Vitro mRNA Transfection Compared to Liposomes.
Authors: Ayad, Camille and Libeau, Pierre and Lacroix-Gimon, Céline and Ladavière, Catherine and Verrier, Bernard
Journal: Pharmaceutics (2021)
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
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
Page updated on October 31, 2024

Ordering information

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50 uL
0.5 mL
5 mL
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Telephone1-800-990-8053
Fax1-800-609-2943
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Physical properties

Solvent

Water

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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 &micro;g of mRNA was transfected with Lipofectamine MessengerMAX and Transfectamine&trade; 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&trade; mRNA Transfection Reagent, most Lipofectamine MessengerMAX transfected samples were scrambled, whereas cells transfected with Transfectamine&trade; mRNA Transfection Reagent looked much healthier (bottom panel).
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 &micro;g of mRNA was transfected with Lipofectamine MessengerMAX and Transfectamine&trade; 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&trade; mRNA Transfection Reagent, most Lipofectamine MessengerMAX transfected samples were scrambled, whereas cells transfected with Transfectamine&trade; mRNA Transfection Reagent looked much healthier (bottom panel).
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 &micro;g of mRNA was transfected with Lipofectamine MessengerMAX and Transfectamine&trade; 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&trade; mRNA Transfection Reagent, most Lipofectamine MessengerMAX transfected samples were scrambled, whereas cells transfected with Transfectamine&trade; mRNA Transfection Reagent looked much healthier (bottom panel).
Cell viability comparison in HeLa cells. HeLa cells were incubated with Transfectamine&trade; 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&trade; Colorimetric WST-8 Cell Quantification Kit (Cat#22770). The higher absorbance at 460 represents more numbers of viable cells.

Documents

pdfSafety data sheet
pdfProduct protocol
pdfCertificate of analysis
Nucleic Acid Building Blocks
Transfection Reagents
Transfectamine™ 6000 CRISPR Transfection Reagent
Transfectamine™ 7000 siRNA Transfection Reagent