相關連結：https://www.miltenyibiotec.com/GB-en/products/macs-cell-cult ...
mRNA 轉染是調節細胞命運同時消除插入到高誘變風險區域的有力工具。使用簡單的基於脂質的轉染試劑，將合成修飾的 mRNA 遞送到細胞中，從而使關鍵發育調節因子、重組酶或標誌物產生瞬時且大量表達。這種開創性的非嵌入式 mRNA 技術徹底改變了體細胞的重編程，並為幹細胞的直接編程和分化提供了新方法。
將成纖維細胞有效重編程為 iPS 細胞
將人成纖維細胞利用 mRNA 重新編程為 iPS 細胞的工作流程。在細胞適應重編程培養基後，每天用轉錄因子的合成修飾 mRNA 轉染細胞 12 天。轉染後 4 小時更換培養基並添加免疫抑製劑 B18R 以抑制先天免疫反應。iPS 細胞集落通常可以在第 14 天檢測到。
使用 mRNA 的轉錄因子表達
我們已經通過免疫細胞化學驗證並建立許多編碼轉錄因子的 mRNA，這些轉錄因子已知可調節細胞命運向神經 (Ascl1 (Mash1)、Lmx1a、Nurr1 (Nr4a2)、Myt1l、NeuroG2、Pou3F2、NeuroD1)、心臟 (Gata4、Mef2c , Mesp1) 和棕色脂肪細胞 (Cebpb, Pparg) 譜系。將原代成纖維細胞接種在 24 孔或 96 孔板中，每 10,000 個細胞轉染 50 至 83 ng mRNA。這些 mRNA 將可用於探索成纖維細胞定向轉化機制。
mRNA相關產品資訊：
| 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 |