Gelatin nanofiber mats with Lipofectamine/plasmid DNA complexes for in vitro genome editing
Gelatin nanofiber mats with Lipofectamine/plasmid DNA complexes for in vitro genome editing
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DOI：
10.1016/j.colsurfb.2022.112561
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发表时间：
2022-05-13
影响因子：
5.8
通讯作者：
Sakai, Shinji
中科院分区：
文献类型：
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作者：
Furuno, Kotoko;Suzuki, Keiichiro;Sakai, Shinji
明胶静电纺纳米纤维垫在生物材料科学中的应用越来越受到关注，如组织工程和药物/基因传递系统。在这项研究中，我们报告了使用电纺丝明胶纳米纤维垫质粒DNA （pDNA）递送。明胶纳米纤维垫与戊二醛交联不溶。在交联垫上，人胚胎肾源性HEK293细胞在培养7天后表现出高活力（>95%），并且在此期间能够增殖。Lipofectamine/pDNA复合物通过溶液浸泡固定在垫子上，HEK293细胞在这些垫子上培养7天表达GFP。此外，HEK293细胞不通过垫释放的pDNA复合物表达GFP，因为将pDNA传递到细胞中的能力丧失了。由于这些垫子可以同时将多种类型的pDNA转染到细胞中，我们已经使用垫子实现了靶向基因组编辑。这些数据强调了含有脂质体/pDNA复合物的明胶纳米纤维垫通过pDNA传递和基因组编辑进行局部基因治疗的潜力。
Gelatin electrospun nanofiber mats are gaining interest for applications in biomaterials science, such as tissue engineering and drug/gene delivery systems. In this study, we report the use of electrospun gelatin nanofiber mats for plasmid DNA (pDNA) delivery. Gelatin nanofiber mats were insolubilized via cross-linking with glutaraldehyde. On the cross-linked mats, human embryonic kidney-derived HEK293 cells demonstrated high viability for 7 days of culture (>95%) and were able to proliferate during that time. The Lipofectamine/pDNA complexes were immobilized on the mats through immersion in a solution, and HEK293 cells cultured on these mats expressed GFP for 7 days. Furthermore, HEK293 cells did not express GFP via the pDNA complexes released from the mats because the ability to deliver pDNA into the cells was lost. Since the mats could be used to transfect multiple types of pDNA into the cells simultaneously, we have achieved targeted genome editing using the mats. These data highlight the potential of gelatin nanofiber mats with Lipofectamine/pDNA complexes for local gene therapy via pDNA delivery as well as genome editing.