Femtosecond optical transfection as a tool for genetic manipulation of human embryonic stem cells
Femtosecond optical transfection as a tool for genetic manipulation of human embryonic stem cells
复制标题
飞秒光转染作为人类胚胎干细胞遗传操作的工具
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
F. Gunn
中科院分区：
文献类型：
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作者：
M. L. Torres;J. Gardner;H. Bradburn;J. King;K. Dholakia;F. Gunn
我们展示了飞秒光转染在人类胚胎干细胞遗传操作中的应用。使用 SLM 与扫描镜相结合的系统，可以快速、有针对性地对单细胞和集落形成的人类胚胎干细胞进行穿孔。在这项工作中，我们展示了使用三剂量的聚焦飞秒激光将带有荧光报告基因的质粒 DNA 成功转染到人类胚胎干细胞中。光穿孔后48小时，大量转染细胞保留了其未分化的形态特征，即大核、高核质比。此外，利用该技术，由不同类型启动子驱动的DNA构建体也成功转染到人类胚胎干细胞中。
We demonstrate the use of femtosecond optical transfection for the genetic manipulation of human embryonic stem cells. Using a system with an SLM combined with a scanning mirror allows poration of both single-cell and colony-formed human embryonic stem cells in a rapid and targeted manner. In this work, we show successful transfection of plasmid DNA tagged with fluorescent reporters into human embryonic stem cells using three doses of focused femtosecond laser. A significant number of transfected cells retained their undifferentiated morphological feature of large nucleus with high nucleus to cytoplasmic ratio, 48h after photoporation. Furthermore, DNA constructs driven by different types of promoters were also successfully transfected into human embryonic stem cells using this technique.
影响因子：
3.5
作者：
Mthunzi, Patience;Dholakia, Kishan;Gunn-Moore, Frank
通讯作者：
Gunn-Moore, Frank