Magnetic tweezers-based 3D microchannel electroporation for high-throughput gene transfection in living cells.
Magnetic tweezers-based 3D microchannel electroporation for high-throughput gene transfection in living cells.
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DOI：
10.1002/smll.201402564
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发表时间：
2015-04-17
期刊：
影响因子：
13.3
通讯作者：
Sooryakumar, Ratnasingham
中科院分区：
文献类型：
--
作者：
Chang, Lingqian;Howdyshell, Marci;Liao, Wei-Ching;Chiang, Chi-Ling;Gallego-Perez, Daniel;Yang, Zhaogang;Lu, Wu;Byrd, John C.;Muthusamy, Natarajan;Lee, L. James;Sooryakumar, Ratnasingham
我们报告了一种基于高通量磁镊的新型 3D 微通道电穿孔系统，能够在单芯片上转染 40,000 个细胞/cm2，用于基因治疗、再生医学和细胞内靶 mRNA 检测，以筛选细胞异质性。单个细胞或单个细胞的有序阵列通过可编程磁场远程引导至穿孔位点，具有高（> 90%）细胞排列效率，使各种转染试剂能够同时递送到细胞中。与传统的基于真空的方法相比，本技术对细胞膜明显更温和，产生> 90%的细胞活力，此外，允许运输转染的细胞以进行进一步分析。说明该系统的多功能性，GATA2分子信标被递送到白血病细胞中以检测与白血病起始相关的GATA2基因的调节水平。 GATA2 阳性细胞和阴性细胞之间的均匀递送和荧光强度的鲜明对比证明了该平台在活细胞中进行基因转移、筛选和检测靶向细胞内标记的关键方面。
We report a novel, high throughput magnetic-tweezers based 3D microchannel electroporation system capable of transfecting 40,000 cells/cm2 on a single-chip for gene therapy, regenerative medicine and intracellular detection of target mRNA for screening cellular heterogeneity. A single cell or an ordered array of individual cells are remotely guided by programmable magnetic fields to poration sites with high (> 90%) cell alignment efficiency to enable various transfection reagents to be delivered simultaneously into the cells. The present technique, in contrast to the conventional vacuum based approach, is significantly gentler on the cellular membrane yielding > 90% cell viability and, moreover, allows transfected cells to be transported for further analysis. Illustrating the versatility of the system, the GATA2 molecular beacon was delivered into leukemia cells to detect the regulation level of the GATA2 gene that is associated with the initiation of leukemia. The uniform delivery and a sharp contrast of fluorescence intensity between GATA2 positive and negative cells demonstrate key aspects of the platform for gene transfer, screening and detection of targeted intracellular markers in living cells.
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