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Messenger RNA (mRNA) is a single-stranded RNA molecule, serving as a transcript of a gene, conveying genetic information from the nucleus to cellular machinery for protein synthesis. Extensively researched in molecular biology and biochemistry, mRNA is now a promising therapeutic agent. In vitro transcribed (IVT) mRNA, synthesized with sequence optimization and chemical modifications, enhances translatability, reduces immunogenicity, and improves stability. Synthetic mRNA offers advantages like the absence of genomic integration risk and cytoplasmic protein expression without nuclear membrane crossing.
Creative Biogene provides a range of premade mRNA products for diverse applications, such as regenerative medicine, vaccination, disease treatment, and cell engineering. Our state-of-the-art facilities and experienced team ensure cost-effective and high-quality solutions tailored to your needs. Contact us for any specific requirements; we look forward to collaborating on your projects.
Key Features of Our Premade mRNA Products
Premade mRNA products find diverse applications across various fields due to their unique properties. In regenerative medicine, these mRNA products serve as efficient tools for directing cellular processes and promoting tissue repair. Vaccination benefits from their use in generating specific immune responses against targeted antigens. In treating hereditary diseases and tumors, premade mRNA allows precise modulation of protein expression, contributing to therapeutic advancements. The absence of genomic integration risk makes them particularly valuable in cell engineering applications, ensuring genetic modifications without unwanted genomic alterations. Specifically, our products can assist you with the following tasks:
Case Study 1
Inducing skeletal muscle cells directly from human pluripotent stem cells (hPSCs) holds promise for in vitro applications such as drug testing, drug discovery, and disease modeling. Researchers employed a groundbreaking method utilizing premade mRNA to rapidly and robustly induce myogenic differentiation of hPSCs. By introducing mRNA encoding MYOD1 and employing siRNA to knock down POU5F1 (OCT4), this integration-free approach yielded functional skeletal myotubes with sarcomere-like structures within days. POU5F1 silencing enhanced MYOD1 recruitment to target promoters, activating significant myogenic gene expression. Transcriptome analyses revealed upregulation of genes associated with IGF- and FGF-signaling, and extracellular matrix, supporting potential applications in regenerative medicine and therapeutic interventions for muscle disorders like muscular dystrophy.
Figure 1. Researchers utilized premade mRNA, synthesized in vitro with enhanced stability, to treat human embryonic stem cells (hESCs) with synMYOD1. The transfection protocol involved multiple applications over several days, leading to sustained POU5F1 expression, as demonstrated by immunostaining for MyHC in synMYOD1-transfected cells. (Akiyama T, et al., 2018)
Case Study 2
Endothelial cells derived from human induced pluripotent stem cells (h-iPSCs) have emerged as a valuable asset in the field of regenerative medicine. Researchers have innovatively employed premade mRNA to establish a rapid, consistent, and highly efficient method for generating h-iPSC–derived endothelial cells (h-iECs). By delivering modified mRNA encoding the transcription factor ETV2 at the intermediate mesodermal stage of differentiation, this approach reproducibly differentiated 13 diverse h-iPSC lines into h-iECs with exceptional efficiency. In contrast, conventional methods relying on endogenous ETV2 were inefficient and notably inconsistent. The resulting h-iECs demonstrated functional competence, including the ability to form perfused vascular networks in vivo. This groundbreaking protocol, critical for vascular therapies, ensures the reliable production of an unlimited number of h-iECs.
Figure 2. Researchers achieved robust endothelial differentiation of h-iPSCs using a two-stage protocol. Stage 1 involved converting h-iPSCs into h-MPCs, and Stage 2 utilized modRNA (ETV2) for the efficient differentiation of h-MPCs into h-iECs. (Wang K, et al., 2020)
Case Study 3
Researchers employed a genetically safe method, in vitro transcribed (IVT) mRNA, to successfully generate induced neural stem cells (iNSCs) from human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs). Transfection with SOX2 mRNA under optimized conditions led to iNSCs with neural stem cell characteristics, including multipotency and self-renewal capacity. Reprogramming of human dermal fibroblasts (HDFs) using SOX2 mRNA demonstrated neural features but with limited proliferation ability compared to embryonic stem cell-derived NSCs. This study showcases UCB-MSCs as a source for direct reprogramming into iNSCs, presenting an integration-free tool with therapeutic potential for neurodegenerative diseases.
Figure 3. Researchers generated SOX2 mRNA, containing the native SOX2 coding sequence, and transfected it into cells. SOX2 gene expression, including both endogenous and exogenous mRNA, increased around 200-fold at 5 days post-induction (DPI). (Kim BE, et al., 2018)
Q: How can premade mRNA products from Creative Biogene be utilized in studying immune response modulation?
A: Premade mRNA products from Creative Biogene can be transfected into immune cells such as dendritic cells or macrophages. For instance, researchers can transfect dendritic cells with mRNA encoding cytokines like IL-34 or chemokines like CCR receptors. The cells can then be stimulated with various antigens or pathogens, and the resulting immune response can be analyzed through cytokine profiling, flow cytometry, or functional assays to understand the modulation of immune responses.
Q: What experimental approach can be employed with premade mRNA products to investigate interferon signaling and antiviral responses?
A: Researchers can transfect cell lines or primary immune cells with premade mRNA encoding interferons such as IFNA4, IFNB1, or IFNG. Subsequently, these cells can be exposed to viral pathogens or stimuli known to induce interferon signaling. The activation of interferon signaling pathways and subsequent antiviral responses can be assessed through gene expression analysis, protein quantification, or functional assays such as viral replication assays or cell viability assays.
Q: How can premade mRNA products aid in the study of cell signaling receptors?
A: Premade mRNA products encoding cell signaling receptors such as TNF receptor family members (e.g., CD40, FAS, TNFRSF1A), cytokine receptors (e.g., IFNLR1, IL1R1), or chemokine receptors (e.g., CCR1-10, CXCR1-6) can be transfected into relevant cell types. After transfection, the cells can be stimulated with specific ligands or cytokines, and downstream signaling events can be analyzed using techniques like Western blotting, immunofluorescence microscopy, or functional assays such as chemotaxis assays or cell proliferation assays.
Q: What experimental design can be implemented to examine transcription factors and regulatory proteins using premade mRNA products?
A: Researchers can transfect cells with premade mRNA encoding transcription factors (e.g., PAX1-9, Mef2c, Tbx5, Gata4) or regulatory proteins involved in apoptosis, cell survival, or gene regulation. Subsequently, the effects of these mRNA products on cellular functions can be studied using techniques such as reporter assays, chromatin immunoprecipitation (ChIP) assays, or cell-based assays assessing apoptosis, proliferation, or differentiation.
Q: How can modified EGFP and Firefly Luciferase mRNA products be utilized to explore innovative mRNA modification and labeling techniques?
A: Modified EGFP and Firefly Luciferase mRNA products can be transfected into cells to study mRNA modification and labeling techniques. Researchers can introduce specific modifications such as 5' cap structures, 3' polyadenylation, or nucleotide substitutions to assess their effects on mRNA stability, translation efficiency, or subcellular localization. Additionally, fluorescently labeled mRNA can be visualized using fluorescence microscopy or flow cytometry to track mRNA uptake, intracellular trafficking, or protein expression dynamics in real-time.
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