Proliferation of genetically modified human cells on electrospun nanofiber scaffolds.
Proliferation of genetically modified human cells on electrospun nanofiber scaffolds.
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DOI：
10.1038/mtna.2012.51
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发表时间：
2012-12-04
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基因编辑是使用单链寡脱氧核苷酸 (ssODN) 在染色体范围内纠正单碱基突变的过程。然而，带有修饰基因的校正细胞的存活和增殖受到一种称为增殖减少表型（RPP）的现象的阻碍。这是实际实施的障碍。为了克服 RPP 问题，我们利用纳米纤维支架作为模板，允许修改后的细胞在基因编辑后恢复、生长和扩展。在这里，我们提出的证据表明，一些带有集成突变增强型绿色荧光蛋白（eGFP）基因并通过基因编辑校正的HCT116-19在聚赖氨酸或纤连蛋白包被的聚己内酯（PCL）纳米纤维支架上增殖。相比之下，来自相同反应方案的细胞在常规培养皿表面和聚赖氨酸（或纤连蛋白）包被的培养皿表面上均没有增殖。因此，在电纺纳米纤维支架上生长转基因（编辑）细胞可促进 RPP 的逆转，并增加基因编辑作为离体基因治疗应用的潜力。
Gene editing is a process by which single base mutations can be corrected, in the context of the chromosome, using single-stranded oligodeoxynucleotides (ssODNs). The survival and proliferation of the corrected cells bearing modified genes, however, are impeded by a phenomenon known as reduced proliferation phenotype (RPP); this is a barrier to practical implementation. To overcome the RPP problem, we utilized nanofiber scaffolds as templates on which modified cells were allowed to recover, grow, and expand after gene editing. Here, we present evidence that some HCT116-19, bearing an integrated, mutated enhanced green fluorescent protein (eGFP) gene and corrected by gene editing, proliferate on polylysine or fibronectin-coated polycaprolactone (PCL) nanofiber scaffolds. In contrast, no cells from the same reaction protocol plated on both regular dish surfaces and polylysine (or fibronectin)-coated dish surfaces proliferate. Therefore, growing genetically modified (edited) cells on electrospun nanofiber scaffolds promotes the reversal of the RPP and increases the potential of gene editing as an ex vivo gene therapy application.
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