Lipofectamine MessengerMAX kit box and one tube with green cap

Superior mRNA transfection efficiency

Invitrogen's Lipofectamine MessengerMAX mRNA Transfection Reagent exhibits up to 5x the efficiency of DNA reagents in neurons and primary cell types.

  • Amazing efficiency for both neuronal transfection and primary cell transfection
  • Fast protein expression with no risk of genomic integration
  • Up to 10x higher cleavage efficiency using mRNA CRISPRs

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Achieve high transfection efficiency in neuronal and primary cells with Lipofectamine MessengerMAX reagent

Lipofectamine MessengerMAX mRNA Transfection Reagent delivers amazing transfection efficiency in neurons and a broad-spectrum of primary cells, enabling improved application outcomes and more biologically relevant research. This reagent’s novel lipid nanoparticle technology is optimized to deliver a high amount of mRNA possible without the nuclear entry step that is required with DNA.

Fluorescent micrographs of neuron, fibroblast, and keratinocyte cells transfected with GFP using MessengerMAX mRNA

Figure 1. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in primary cells (fresh isolated mouse cortical neurons, BJ fibroblasts, and primary keratinocytes). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (500 ng/well) in a 24-well format. The leading DNA delivery reagent was used to deliver GFP DNA (500 ng/well), and GFP was analyzed 24 hours post-transfection.

Fluorescent micrographs of stem cell lines iPSCs, H9 ESCs, hNSCs, and mESCs transfected with GFP using MessengerMAX mRNA

Figure 2. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in various stem cells (Gibco iPSCs, H9 ESCs, mESCs, hNSCs). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (250 ng/well) in a 48-well format. The leading DNA delivery reagent was used to deliver GFP DNA (250 ng/well), and GFP was analyzed 24 hours post-transfection.

Fluorescent micrographs of bend3, SH-SY-SY, and SK-N-SH cancer cells transfected with GFP using MessengerMAX mRNA

Figure 3. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in various cell lines that are very difficult to transfect (bEnd.3, SH-SY5Y, SK-N-SH, A431, RAW 264.7, and HT29). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (500 ng/well) in a 24-well format. The leading DNA delivery reagent was used to deliver GFP DNA (500 ng/well), and GFP was analyzed 24 hours post-transfection.

Fluorescent micrographs of neuron, fibroblast, and keratinocyte cells transfected with GFP using MessengerMAX mRNA

Figure 1. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in primary cells (fresh isolated mouse cortical neurons, BJ fibroblasts, and primary keratinocytes). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (500 ng/well) in a 24-well format. The leading DNA delivery reagent was used to deliver GFP DNA (500 ng/well), and GFP was analyzed 24 hours post-transfection.

Fluorescent micrographs of stem cell lines iPSCs, H9 ESCs, hNSCs, and mESCs transfected with GFP using MessengerMAX mRNA

Figure 2. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in various stem cells (Gibco iPSCs, H9 ESCs, mESCs, hNSCs). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (250 ng/well) in a 48-well format. The leading DNA delivery reagent was used to deliver GFP DNA (250 ng/well), and GFP was analyzed 24 hours post-transfection.

Fluorescent micrographs of bend3, SH-SY-SY, and SK-N-SH cancer cells transfected with GFP using MessengerMAX mRNA

Figure 3. Lipofectamine MessengerMAX outperforms leading DNA delivery reagent and leading mRNA delivery reagent in various cell lines that are very difficult to transfect (bEnd.3, SH-SY5Y, SK-N-SH, A431, RAW 264.7, and HT29). Lipofectamine MessengerMAX and the leading mRNA delivery reagent were used to deliver GFP mRNA (500 ng/well) in a 24-well format. The leading DNA delivery reagent was used to deliver GFP DNA (500 ng/well), and GFP was analyzed 24 hours post-transfection.


mRNA transfection: Fast protein expression with no risk of genomic integration

While DNA transfection is the most common method used to express a gene of interest, this process involves several steps, including DNA entry into the nucleus. The entry of DNA into the nucleus is often one of the most difficult transfection steps, especially in hard-to-transfect cells.

In contrast, mRNA transfection is simpler than DNA transfection as the mRNA transcript can be delivered directly into the cells and does not require nuclear entry (step 4, Figure 4) for expression. In addition, mRNA transfection is cell cycle independent and helps eliminate the risk of genomic integration.

Lipofectamine MessengerMAX Transfection Reagent, specifically designed for mRNA transfection, results in fast protein expression with homogeneity of expression among the transfected cells.

Diagram showing the steps of DNA and mRNA transfection from the reagent-DNA or -mRNA complex to the protein

Figure 4. DNA vs. mRNA transfection.


Up to 10X higher cleavage efficiency using CRISPR mRNAs

Lipofectamine MessengerMAX reagent increases the likelihood of cleavage and recombination with CRISPR nuclease mRNA through highly efficient transfection, helping maximize the efficiency of CRISPR-driven genetic modifications and simplifying downstream processes.

Bar graph showing that Cas9 mRNA and IVT gRNA transfected using Lipofectamine MessengerMAX showed higher cleavage efficiency than those transfected with competitor products. The all-in-one CRISPR plasmid transfected with Lipofectamine 3000 also showed much lower cleavage efficiency.

Figure 5. Cleavage efficiency of various Invitrogen GeneArt CRISPR formats targeting the HPRT locus in Gibco iPS cells in a 12-well format. Invitrogen Lipofectamine 3000 reagent was used to deliver CRISPR nuclease all-in-one plasmid DNA; Lipofectamine MessengerMAX reagent and two leading mRNA delivery reagents were used to deliver an all-RNA CRISPR format (Cas9 mRNA + IVT gRNA). Cleavage efficiency was determined using the Invitrogen GeneArt Genomic Cleavage Detection Kit 72 hours post-transfection.


Getting started with mRNA delivery in cells is as easy as 1-2-3!

Prepare DNA template

Prepare your DNA template with a T7 promoter. You may choose to clone your gene with an Invitrogen Gateway pcDNA-DEST40 Vector or amplify it with PCR and T7-containing primers.

Preparing DNA template with a T7 promoter

Generate mRNA

Transcribe your template DNA with the Invitrogen mMESSAGE mMACHINE T7 ULTRA Transcription Kit to generate mRNA for transfection.

Invitrogen mMESSAGEm MACHINE T7 ULTRA Transcription Kit

Transfect mRNA

Transfect your mRNA with our simple Lipofectamine MessengerMAX transfection protocol.
 Download the Lipofectamine MessengerMAX protocol

mRNA workflow

Prepare DNA template

Prepare your DNA template with a T7 promoter. You may choose to clone your gene with an Invitrogen Gateway pcDNA-DEST40 Vector or amplify it with PCR and T7-containing primers.

Preparing DNA template with a T7 promoter

Generate mRNA

Transcribe your template DNA with the Invitrogen mMESSAGE mMACHINE T7 ULTRA Transcription Kit to generate mRNA for transfection.

Invitrogen mMESSAGEm MACHINE T7 ULTRA Transcription Kit

Transfect mRNA

Transfect your mRNA with our simple Lipofectamine MessengerMAX transfection protocol.
 Download the Lipofectamine MessengerMAX protocol

mRNA workflow
Cell typeLipofectamine MessengerMAX transfection efficiency (%)
MDA-MB-231
A431
A549
bEnd.3
BJ fibroblast
H9 ESCs
Primary hepatocytes
Hep G2
HT-29
iPSCs
Primary keratinocytes
L929
LNCaP
hNSCs
MCF7
Neuro-2a
Primary neurons
RAW 264.7
RBL
SK-N-SH
SH-SY5Y

Transfection efficiency (%): <30%     30–50%    51–79%    >80%



Frequently asked questions (FAQs) about working with mRNA

Why is mRNA unstable?

Is it difficult to work with mRNA?

Do mRNA transfections take longer than DNA transfections?

Are mRNA transfections more expensive?
 

How to get started with Lipofectamine MessengerMAX reagent

To help ensure validity of transfection results, we strongly recommend the use of a positive control mRNA, available from various third-party vendors, including TriLink BioTechnologies (e.g., EGFP mRNA).

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Packaging must protect the product inside—but that doesn’t have to mean sacrificing sustainability. Lipofectamine products are shipped at ambient temperature, diverting 24,000 ft3 of Expanded Polystyrene (EPS) coolers from landfills annually. We reduce the carbon footprint from packaging and shipping these products by 80 tons (CO2 equivalents).

Learn more about sustainable solutions
 Download the green fact sheet

Resources

Our Lipofectamine reagent-specific protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. Search this library to find protocols and citations curated to fit your experimental needs. Filter by cell type, payload, and product of choice to easily unlock your answers.

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Transfection FAQs

Search our extensive collection of Transfection FAQs to find the answers related to Lipofectamine MessengerMAX reagent.

For Research Use Only. Not for use in diagnostic procedures.

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