Jérôme Valérie, Synatschke Christopher V, Freitag Ruth
Process Biotechnology, University of Bayreuth, 95440 Bayreuth, Germany.
Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
ACS Omega. 2020 Oct 5;5(41):26640-26654. doi: 10.1021/acsomega.0c03367. eCollection 2020 Oct 20.
Nonviral DNA vectors are promising alternatives to viral ones. Their use in DNA medicine is limited by an inability to transfect, for example, nondividing or suspension cells. In recent years, star-shaped synthetic polycationic vectors, so called "Nanostars", have shown some promise in this regard, at least when compared to the "gold standard" in nonviral vectors, namely, linear poly(ethyleneimine) (l-PEI). It has been hypothesized that an ability to transiently destabilize cellular membranes is partially responsible for the phenomenon. This hypothesis is investigated here, taking human leukemia suspension cells (Jurkat cells) as an example. Contrary to l-PEI, the Nanostars promote the cellular uptake of small, normally membrane-impermeant molecules (trypan blue and propidium iodide) as well as that of fluorescent polystyrene beads (average diameter 100 nm). Since Nanostars, but not l-PEI, are apparently able to deliver DNA to nuclei of nondividing cells, nuclear uptake is, in addition, investigated with isolated cell nuclei. Our results provide evidence that Nanostars are more efficient than l-PEI in increasing the nuclear membrane association/permeability, allowing accumulation of their cargo on/in the nucleus.
非病毒DNA载体是病毒载体很有前景的替代物。它们在DNA医学中的应用受到限制,例如无法转染非分裂细胞或悬浮细胞。近年来,星形合成聚阳离子载体,即所谓的“纳米星”,在这方面已显示出一些前景,至少与非病毒载体的“金标准”,即线性聚(乙烯亚胺)(l-PEI)相比是这样。据推测,瞬时破坏细胞膜的能力部分导致了这一现象。本文以人白血病悬浮细胞(Jurkat细胞)为例对这一假设进行了研究。与l-PEI相反,纳米星促进了小的、通常不能透过细胞膜的分子(台盼蓝和碘化丙啶)以及荧光聚苯乙烯珠(平均直径100nm)的细胞摄取。由于纳米星显然能够将DNA递送至非分裂细胞的细胞核,而l-PEI则不能,因此,我们还利用分离的细胞核研究了核摄取情况。我们的结果提供了证据,表明纳米星在增加核膜结合/通透性方面比l-PEI更有效,从而使它们携带的物质能够在细胞核上/内积累。