Mesenchymal stem cells (MSCs) have been intensively investigated and widely applied in regenerative medicine and immune modulation. However, their efficacy declines during the aging or disease process. Thus, genome-edited MSCs with over-expression or inhibition of specific genes hold a great deal of promise in terms of their therapeutic application. Here we optimized the direct PCR approach for rapid identification of genome-edited MSCs with only ten cells required, which reduces the time and labor to expand the MSC colonies. Combined with our previously optimized guide RNA structure and plasmid construction strategy for Cas9, we successfully identified MSC colonies over-expressing IL-10 in the AAVS1 locus.

中文翻译：

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通过 Cas9 快速鉴定基因组编辑的间充质干细胞集落。


间充质干细胞（MSC）已被深入研究并广泛应用于再生医学和免疫调节。然而，它们的功效在衰老或疾病过程中会下降。因此，过度表达或抑制特定基因的基因组编辑的 MSC 在其治疗应用方面具有很大的前景。在这里，我们优化了直接 PCR 方法，只需 10 个细胞即可快速鉴定基因组编辑的 MSC，从而减少了扩展 MSC 集落的时间和劳动力。结合我们之前优化的Cas9引导RNA结构和质粒构建策略，我们成功鉴定了在AAVS1位点过表达IL-10的MSC集落。