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Real-time intracellular transport of gene nanocarriers studied by multiple particle tracking

机译:通过多粒子跟踪研究基因纳米载体的实时细胞内转运

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We used real-time multiple particle tracking to quantitatively characterize the type and rates of transport of gene nanocarriers within live cells. The heterogeneous cytoplasmic transport of polyethylenimine (PEI)/DNA gene carriers was quantified by tracking their mean-square displacements over time and classified into active and nonactive transport populations on the basis of their effective diffusivities versus time. Nonactive gene carriers frequently displayed hop-diffusion trajectories, suggesting a porous cytoplasmic network of flexible biopolymers or sequential attachment and detachment events. Microtubule-dependent active transport of gene carriers resulted in an effective diffusivity 30-fold greater than that of nonactive carriers (at a time scale of 3 s). Compared to nonactive carriers in control cells with intact microtubules, microtubule depolymerization enhanced short-range motion of gene carriers but resulted in similar long-range transport. Multiple particle tracking characterizes gene carrier transport in complex biological environments and, therefore, may be a useful tool in quantifying rate-limiting steps in gene delivery within cells and other biological media.
机译:我们使用实时多粒子跟踪来定量表征活细胞内基因纳米载体的运输类型和速率。聚乙烯亚胺(PEI)/ DNA基因载体的异质胞质运输通过跟踪其随时间的均方位移进行定量,并根据有效扩散率与时间的关系分为活跃和不活跃的运输种群。非活性基因携带者经常表现出跳跃扩散的轨迹,这表明柔性生物聚合物的多孔细胞质网络或顺序的附着和脱离事件。基因载体的微管依赖性主动转运导致有效扩散率是非活性载体的有效扩散率的30倍(在3 s的时间尺度上)。与具有完整微管的对照细胞中的非活性载体相比,微管解聚增强了基因载体的短程运动,但导致了类似的长程运输。多重粒子跟踪表征了复杂生物环境中基因载体的运输,因此,可能是量化细胞和其他生物介质中基因传递的限速步骤的有用工具。

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