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Improved dynamic monitoring of transcriptional activity during longitudinal analysis in the mouse brain

机译:改进的动态监测小鼠大脑纵向分析过程中的转录活性

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Bioluminescence imaging has proven to be a highly sensitive technique for assessingin vitrotranscriptional activity toward understanding gene regulation patterns; however, application of this technique is limited for brain research. In particular, the poor spatiotemporal resolution is a major hurdle for monitoring the dynamic changes of transcriptional activity in specific regions of the brain during longitudinal analysis of living animals. To overcome this limitation, in this study, we modified a lentivirus-based luciferase glucocorticoid receptor (GR) reporter by inserting destabilizing sequence genes, and then the reporter was stereotaxically injected in the mouse infralimbic prefrontal cortex (IL-PFC). Using this strategy, we could successfully pin-point and monitor the dynamic changes in GR activity in IL-PFC during normal stress adaptation. The modified reporter showed a 1.5-fold increase in temporal resolution for monitoring GR activity compared to the control, with respect to the intra-individual coefficients of variation. This novelin vivomethod has broad applications, as it is readily adaptable to different types of transcription factor arrays as well spanning wide target regions of the brain to other organs and tissues.
机译:已经证明,生物发光成像是一种用于评估体外转录活性以了解基因调控模式的高度灵敏的技术。但是,该技术的应用仅限于大脑研究。特别地,较差的时空分辨率是在对活体动物进行纵向分析期间监测大脑特定区域转录活性动态变化的主要障碍。为了克服此限制,在这项研究中,我们通过插入去稳定序列基因来修饰基于慢病毒的萤光素酶糖皮质激素受体(GR)报告基因,然后将报告基因立体定位于小鼠下肢前额叶皮层(IL-PFC)。使用此策略,我们可以在正常压力适应过程中成功查明并监测IL-PFC中GR活性的动态变化。相对于个体内部变异系数,与对照组相比,改良的报告基因显示出用于监测GR活动的时间分辨率提高了1.5倍。这种新颖的体内方法具有广泛的应用前景,因为它很容易适应不同类型的转录因子阵列,并且将大脑的宽靶区域跨越到其他器官和组织。

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