MS-1 magA: Revisiting Its Efficacy as a Reporter Gene for MRI.
MS-1 magA: Revisiting Its Efficacy as a Reporter Gene for MRI.
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DOI：
10.1177/1536012116641533
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发表时间：
2016
影响因子：
2.8
通讯作者：
Taylor A
中科院分区：
文献类型：
--
作者：
Pereira SM;Williams SR;Murray P;Taylor A
最近，参与趋磁细菌中磁性纳米颗粒生物矿化的细菌基因被提出作为磁共振成像(MRI)的报告程序。在这样的系统中，细菌基因在哺乳动物细胞中的表达据称会导致细胞内更高浓度的铁或氧化铁的生物矿化，从而导致松弛速率的增强，这一点可以通过MRI检测到。在这里，我们发现磁性磁螺菌MAGA基因的结构性表达对人胚胎肾脏(HEK)细胞是耐受的，但在小鼠间充质/基质细胞和肾源性干细胞中诱导了强烈的毒性作用，严重限制了其作为干细胞报告基因的有效利用。尽管MAGA被认为与铁的转运有关，但当它在HEK细胞中表达时，并不影响与铁稳态相关的内源性基因的转录。此外，MAGA诱导HEK细胞铁摄取的增加并不显著，这表明即使对于能够耐受其表达的细胞类型来说，该基因也是一个很差的报告基因。我们建议应谨慎使用MAGA作为干细胞，其作为报告基因的有效性需要在逐个细胞的基础上进行仔细的评估。
Bacterial genes involved in the biomineralization of magnetic nanoparticles in magnetotactic bacteria have recently been proposed as reporters for magnetic resonance imaging (MRI). In such systems, the expression of the bacterial genes in mammalian cells purportedly leads to greater concentrations of intracellular iron or the biomineralization of iron oxides, thus leading to an enhancement in relaxation rate that is detectable via MRI. Here, we show that the constitutive expression of the magA gene from Magnetospirillum magnetotacticum is tolerated by human embryonic kidney (HEK) cells but induces a strong toxic effect in murine mesenchymal/stromal cells and kidney-derived stem cells, severely restricting its effective use as a reporter gene for stem cells. Although it has been suggested that magA is involved in iron transport, when expressed in HEK cells, it does not affect the transcription of endogenous genes related to iron homeostasis. Furthermore, the magA-induced enhancement in iron uptake in HEK cells is insignificant, suggesting this gene is a poor reporter even for cell types that can tolerate its expression. We suggest that the use of magA for stem cells should be approached with caution, and its efficacy as a reporter gene requires a careful assessment on a cell-by-cell basis.