Selective gene transfection of individual cells in vitro with plasmonic nanobubbles.
Selective gene transfection of individual cells in vitro with plasmonic nanobubbles.
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DOI：
10.1016/j.jconrel.2011.02.006
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发表时间：
2011-06-10
期刊：
影响因子：
--
通讯作者：
Lapotko DO
中科院分区：
文献类型：
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作者：
Lukianova-Hleb EY;Samaniego AP;Wen J;Metelitsa LS;Chang CC;Lapotko DO
真核细胞的基因递送和转染广泛用于研究和开发基因细胞疗法。然而，当必须处理异质细胞系统时，现有方法缺乏选择性、有效性和安全性。我们报告了一种利用等离子体纳米气泡（PNB）进行递送和转染的新方法。 PNB 是一种新颖的可调谐细胞制剂，具有双重机械和光学作用，这是由于在暴露于激光脉冲时在瞬时加热的金纳米颗粒周围形成蒸汽纳米气泡。 PNB 能够将细胞外 cDNA 质粒机械注射到单个目标活细胞、培养的白血病细胞和人 CD34+CD117+ 干细胞的细胞质中，并在这些细胞中表达绿色荧光蛋白 (GFP)。 PNB 的产生和寿命与 PNB 处理的细胞中绿色荧光蛋白的表达相关。 PNB 的光散射还提供了靶细胞的检测以及单细胞水平 cDNA 注射的指导。在两种细胞模型中，PNB 在单次脉冲治疗中均表现出基因转染效果，具有高选择性、有效性和安全性。因此，PNB 在单脉冲过程中提供单细胞水平的靶向基因递送，可用于安全有效的基因治疗。
Gene delivery and transfection of eukaryotic cells is widely used for research and for developing gene cell therapy. However, the existing methods lack selectivity, efficacy and safety when heterogeneous cell systems must be treated. We report a new method that employs plasmonic nanobubbles (PNBs) for delivery and transfection. A PNB is a novel, tunable cellular agent with a dual mechanical and optical action due to the formation of the vapor nanobubble around a transiently heated gold nanoparticle upon its exposure to a laser pulse. PNBs enabled the mechanical injection of the extracellular cDNA plasmid into the cytoplasm of individual target living cells, cultured leukemia cells and human CD34+CD117+ stem cells and expression of a green fluorescent protein (GFP) in those cells. PNB generation and lifetime correlated with the expression of green fluorescent protein in PNB-treated cells. Optical scattering by PNBs additionally provided the detection of the target cells and the guidance of cDNA injection at single cell level. In both cell models PNBs demonstrated a gene transfection effect in a single pulse treatment with high selectivity, efficacy and safety. Thus, PNBs provided targeted gene delivery at the single cell level in a single pulse procedure that can be used for safe and effective gene therapy.