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mRNA transfection for stem cell differentiation and reprogramming
mRNA transfection is a powerful tool for modulating cell fate while eliminating the risk of insertional mutagenesis. Using simple lipid-based transfection, modified mRNAs are delivered into the cell allowing high-level, transient expression of key developmental regulators, recombinases, or markers. This pioneering non-integrative mRNA technology has revolutionized reprogramming of somatic cells and offers new approaches for direct programming and differentiation of stem cells.
Efficient reprogramming of fibroblasts into iPS cells
Workflow for efficient reprogramming of human fibroblasts into iPS cells. After adaption of the cells to the reprogramming medium, cells are transfected daily for 12 days with modified mRNAs coding for reprogramming transcription factors. Medium is replaced 4 h after transfection and immune suppressor B18R is added to inhibit innate immune responses. iPS cell colonies can be detected usually at day 14.
mRNA-based expression of transcription factors
We have set up and validated by immunocytochemistry a number of mRNAs coding for transcription factors known to modulate cell fate towards neural (Ascl1 (Mash1), Lmx1a, Nurr1 (Nr4a2), Myt1l, NeuroG2, Pou3F2, NeuroD1), cardiac (Gata4, Mef2c, Mesp1) and brown adipocyte (Cebpb, Pparg) lineages. Primary fibroblasts were seeded in 24-well or 96-well plates and transfected with 50 to 83 ng mRNA per 10,000 cells. These RNAs will be used to explore the RNA technology for directed conversion of fibroblasts.
Related products:
| Research Area | Order no. | Products | Size | Remarks | Application |
| CRISPR-Cas9 | 130-107-751 | StemMACS™ Cas9 Nuclease mRNA | 20 µg | encodes the Cas9 nuclease from Streptomyces pyogenes | site-directed single strand breaks within the genomic target DNA |
| CRISPR-Cas9 | 130-107-679 | StemMACS™ Cas9 Nickase mRNA | 20 µg | encodes a D10A mutated version of Cas9 from Streptomyces pyogenes | only the strand of the DNA that is complementary to the guide RNA sequence is cleaved |
| Cre-LoxP recombination | 130-101-113 | StemMACS™ Cre Recombinase mRNA | 20 µg | encodes a codon-optimized variant of Cre recombinase | site-specific recombination between loxP sites |
| Flp-FRT recombination | 130-106-769 | StemMACS™ Flp Recombinase mRNA | 20 µg | encodes a codon optimized version of Flp recombinase. | recombination between two identical flippase recognition target (FRT) sites |
| fluorescent protein | 130-120-971 | StemMACS™ YPet mRNA | 20 µg | encodes the yellow fluorescent protein YPet | Positive control for mRNA transfections Optimization of mRNA transfection protocols Transient labeling of cells by transfection |
| fluorescent protein | 130-120-975 | StemMACS™ mCherry mRNA | 20 µg | encodes the red fluorescent protein mCherry | Positive control for mRNA transfections Optimization of mRNA transfection protocols Transient labeling of cells by transfection |
| fluorescent protein | 130-101-114 | StemMACS™ eGFP mRNA | 20 µg | enhanced green fluorescent protein (eGFP) | Positive control for mRNA transfections Optimization of mRNA transfection protocols Transient labeling of cells by transfection |
| fluorescent protein | 130-101-119 | StemMACS™ Nuclear eGFP mRNA | 20 µg | encodes an enhanced green fluorescent protein (eGFP) linked to a nuclear localization signal | Positive control for mRNA transfections Optimization of mRNA transfection protocols Transient labeling of cells by transfection |
| iPSC reprogramming | 130-101-075 | StemMACS™ Sox2 mRNA, human | 20 µg | encodes the human transcription factor Sox2 | Modulation of pluripotency Reprogramming |
| iPSC reprogramming | 130-101-112 | StemMACS™ c-Myc mRNA, human | 20 µg | encodes a stabilized version of the transcription factor c-Myc | Modulation of pluripotency Reprogramming |
| iPSC reprogramming | 130-101-115 | StemMACS™ Klf4 mRNA, human | 20 µg | encodes the transcription factor Krüppel-like factor 4 (Klf4) | Modulation of pluripotency Reprogramming |
| iPSC reprogramming | 130-101-117 | StemMACS™ Lin28 mRNA, human | 20 µg | encodes the human RNA-binding protein Lin28 | Modulation of pluripotency Reprogramming |
| iPSC reprogramming | 130-101-118 | StemMACS™ Nanog mRNA, human | 20 µg | encodes the human transcription factor Nanog | Modulation of pluripotency Reprogramming |
| Neuronal reprogramming | 130-104-369 | StemMACS™ Mash1 (Ascl1) mRNA, human | 20 µg | encodes the human transcription factor Mash1 | Neuronal differentiation of pluripotent stem cells Conversion of fibroblasts into induced neuronal (iN) cells |
| Neuronal reprogramming | 130-104-370 | StemMACS™ Brn2 mRNA, human | 20 µg | encodes the human transcription factor Brn2 | Neuronal differentiation of pluripotent stem cells Conversion of fibroblasts into induced neuronal (iN) cells |
| Neuronal reprogramming | 130-104-379 | StemMACS™ Myt1l mRNA, human | 20 µg | encoding the human transcription factor Myt1l | Neuronal differentiation of pluripotent stem cells Conversion of fibroblasts into induced neuronal (iN) cells |
| Neuronal reprogramming | 130-104-382 | StemMACS™ NeuroD1 mRNA, human | 20 µg | encodes the neural transcription factor NeuroD1 | Neuronal differentiation of pluripotent stem cells Conversion of fibroblasts into induced neuronal (iN) cells |
| Neuronal reprogramming | 130-104-383 | StemMACS™ NeuroG2 mRNA, human | 20 µg | encodes the neural transcription factor NeuroD1 | Neuronal differentiation of pluripotent stem cells Conversion of fibroblasts into induced neuronal (iN) cells |
| dopaminergic neurons differentiation | 130-104-381 | StemMACS™ Lmx1a mRNA, human | 20 µg | encodes the human transcription factor Lmx1a | Differentiation of pluripotent stem cells into dopaminergic neurons |
| dopaminergic neurons differentiation | 130-104-384 | StemMACS™ Nurr1 (NR4a2) mRNA, human | 20 µg | the human transcription factor Nurr1 | Differentiation of pluripotent stem cells into dopaminergic neurons |
| adipocyte differentiation | 130-104-386 | StemMACS™ Pparg mRNA, human | 20 µg | encodes the human transcription factor PPAR-gamma | differentiation into adipose tissue |
| adipocyte differentiation | 130-104-377 | StemMACS™ Cebpb mRNA, human | 20 µg | encodes the transcription factor C/EBP-beta | differentiation into adipose tissue |
| lipid-based transfection system | 130-104-463 | StemMACS™ mRNA Transfection Kit | 0.5 mL | for efficient mRNA delivery into various cell types |