Introducing the mRNA transfection reagent with up to 5X the efficiency of DNA reagents in neurons and primary cell types.
Featured application note: Neurobiology transfection guide
The field of neurobiology is rapidly growing, with researchers providing new insights into a variety of areas essential for human health, including memory, mood disorders, aging, and disease. As the field moves toward more physiologically relevant models, such as primary neurons and small animal models, the need for new nucleic acid delivery tools is rapidly increasing. Download this guide to assist you in this effort.
"I tested the MessengerMAX on rat pancreatic primary alpha and beta cells, using a positive control eGFP. The expression worked perfectly. I then checked my target gene expression and it increased by more than 1000 fold!" (see data) Dr Mounia Heddad-Masson, Hospital of Geneva (diabetology dpt.)
"Lipofectamine MessengerMAX reagent can transfect mesenchymal stem cells at high efficiency wiithout the toxic effects of viral vectors." Dr. Daniel Cohen, University of Pennsylvania
In order to generate your mRNA transcript, we recommend the following transcription kit:
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).
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. That's because our novel lipid nanoparticle technology is optimized to deliver the highest amount of mRNA possible without the nuclear entry step that is required with DNA.
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 posttransfection.
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 posttransfection.
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 posttransfection.
| Cell type | Lipofectamine® 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%
Transfection of mRNA with Lipofectamine MessengerMAX typically results in faster protein expression with great homogeneity of expression among the transfected cells. Additionally, delivery of mRNA does not require nuclear entry (step 4), which eliminates the risk of genomic integration and transfection efficiency becomes cell cycle independent.
Figure 4. DNA vs. mRNA transfection.
1 |
Prepare DNA templatePrepare 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.
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2 |
Generate mRNATranscribe your template DNA with the Invitrogen™ mMESSAGE mMACHINE T7 ULTRA Transcription Kit to generate mRNA for transfection.
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3 |
Transfect mRNATransfect your mRNA with our simple Lipofectamine MessengerMAX transfection protocol. Download the Lipofectamine MessengerMAX protocol
Figure 5. mRNA workflow |
Lipofectamine MessengerMAX reagent increases the likelihood of cleavage and recombination with Invitrogen™ GeneArt™ CRISPR Nuclease mRNA through highly efficient transfection, maximizing the efficiency of CRISPR-driven genetic modifications and simplifying downstream processes.
Figure 6. Cleavage efficiency of various GeneArt™ CRISPR formats targeting the HPRT locus in Gibco™ iPS cells in a 12-well format. Lipofectamine™ 3000 reagent was used to deliver CRISPR nuclease all-in-one plasmid DNA; Lipofectamine MessengerMAX 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 GeneArt™ Genomic Cleavage Detection Kit 72 hours post-transfection.
For Research Use Only. Not for use in diagnostic procedures.
