Gene-activated matrices (GAMs) encoding pivotal transcription factors (TFs) represent a powerful tool to direct stem cell specification for tissue engineering applications. However, current TF-based GAMs activated with pDNA, are challenged by their low transfection efficiency and delayed transgene expression. Here, we report a GAM technology activated with mRNAs encoding TFs SOX9 (cartilage) and MYOD (muscle). We find that these mRNA-GAMs induce a higher and faster TF expression compared to pDNA-GAMs, especially in the case of RNase resistant mRNA sequences. This potent TF expression was translated into a high synthesis of cartilage- and muscle-specific markers, and ultimately, into successful tissue specification in vitro. Additionally, we show that the expression of tissue-specific markers can be further modulated by altering the properties of the mRNA-GAM environment. These results highlight the value of this GAM technology for priming cell lineage specification, a key centerpiece for future tissue engineering devices.

编码关键转录因子（TFs）的基因激活基质（GAM）代表了指导干细胞规范用于组织工程应用的强大工具。但是，当前的被pDNA激活的基于TF的GAM受到其低转染效率和延迟的转基因表达的挑战。在这里，我们报告了一种GAM技术，该技术被编码TFs SOX9（软骨）和MYOD（肌肉）的mRNA激活。我们发现与pDNA-GAM相比，这些mRNA-GAM诱导更高和更快的TF表达，尤其是在RNase抗性mRNA序列的情况下。这种有效的TF表达被转化为软骨和肌肉特异性标记物的高度合成，并最终转化为成功的体外组织规格。另外，我们表明，可以通过改变mRNA-GAM环境的特性来进一步调节组织特异性标志物的表达。这些结果凸显了该GAM技术对于启动细胞谱系规范的价值，而该谱系规范是未来组织工程设备的关键核心。