Secreted luciferase for in vivo evaluation of systemic protein delivery in mice.
Secreted luciferase for in vivo evaluation of systemic protein delivery in mice.
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分泌型荧光素酶，用于体内评估小鼠全身蛋白质递送。
DOI：
10.1007/s12033-012-9519-6
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发表时间：
2013-01
影响因子：
2.6
通讯作者：
Pan D
中科院分区：
文献类型：
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作者：
El-Amouri SS;Cao P;Miao C;Pan D
天然分泌的Gaussia荧光素酶（Gluc）已被用作生物发光成像（BLI）评价的报告基因。然而，Gluc用于体内监测全身蛋白递送及其自然生物分布的潜在应用尚未研究。为了检查Gluc分泌和摄取曲线，我们通过流体动力学尾静脉注射将Glu c编码质粒注射到小鼠中。全身BLI显示在爪垫获得的成像定量与血液Gluc活性直接相关。当通过使用肝启动子将基因表达限制在肝脏时，体内Gluc生物分布分析显示肾/膀胱、胃/肠和肺为主要摄取器官。三维BLI确定肝/胃和肺作为主要的内部发光源，证明了通过3D BLI在循环中具有高背景信号的情况下检测活体动物中的主要摄取器官的可行性。值得注意的是，Gluc注射小鼠毛细血管耗竭脑样品中的Gluc水平与对照组相当，表明Gluc可能无法穿过血脑屏障。在各种类型的细胞系中评估了Gluc摄取动力学和细胞内半衰期，暗示了非特异性胞饮作用的参与。这些结果表明，基于葡萄糖的系统可以提供一个有用的工具，在体内评价蛋白质/药物的生物分布后，全身输送。
A naturally secreted Gaussia luciferase (Gluc) has been utilized as a reporter for bioluminescence imaging (BLI) evaluation. However, the potential application of Gluc for in vivo monitoring of systemic protein delivery, as well as its natural biodistribution, has not been studied. To examine Gluc secretion and uptake profile, we injected Gluc-encoding plasmids into mice by hydrodynamic tail-vein injection. Whole-body BLI showed that imaging quantification obtained at pawpad was directly correlated to blood Gluc activities. When gene expression was restricted to the liver by the use of a hepatic promoter, in vivo Gluc biodistribution analysis revealed the kidney/bladder, stomach/intestine and lung as the major uptake organs. Three-dimensional BLI identified liver/stomach and lung as the main internal luminescent sources, demonstrating the feasibility of detecting major uptake organs in live animals by 3D BLI with high background signals in circulation. Notably, Gluc levels in capillary-depleted brain samples from Gluc-injected mice were comparable to controls, suggesting Gluc may not cross the blood-brain barrier. Gluc uptake kinetics and intracellular half-life were assessed in various types of cell lines, implicating the involvement of non-specific pinocytosis. These results suggest that Gluc-based system may provide a useful tool for in vivo evaluation of protein/agent biodistribution following systemic delivery.
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蛋白水解活性的增加是表达中间 t-癌基因的内皮细胞形态发生行为异常的原因
影响因子：
64.5
作者：
MONTESANO, R;PEPPER, MS;ORCI, L
通讯作者：
ORCI, L
DOI：
10.1073/pnas.0701185104
发表时间：
2007-04-24
影响因子：
11.1
作者：
Hewett, Jeffrey W.;Tannous, Bakhos;Breakefield, Xandra O.
通讯作者：
Breakefield, Xandra O.
影响因子：
11.4
作者：
Giodini, Alessandra;Cresswell, Peter
通讯作者：
Cresswell, Peter
DOI：
10.1016/j.bbrc.2005.11.063
发表时间：
2006-01-13
影响因子：
3.1
作者：
Al-Dosari, M;Zhang, GS;Liu, DX
通讯作者：
Liu, DX
影响因子：
82.9
作者：
通讯作者：
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