Understanding Cell Division

总结 此设备申请补充的母公司补助金（1 R35 GM 126980）有权 “了解细胞分裂”该奖项涵盖了主要的研究重点， Gorbsky实验室，分析了染色体运动的机制， 有丝分裂细胞周期，并表征的因素和条件， 染色体不稳定性。我们的大部分研究涉及高分辨率成像， 表达荧光标记蛋白质的培养细胞，并用RNA操作 干扰或CRISPR基因敲除和突变。Mizar光片提供了 具有最小光毒性的高分辨率成像。在实验室中使用的方法， 询问蛋白质功能包括siRNA或敲除降解决定子/GFP标签进入内源性 基因.我们还使用“修剪”技术，通过这种技术， 抗体和Trim 13 E3泛素连接酶，显微注射到活细胞中， 降解内源性蛋白质而无需遗传操作。请求的 升级到我们的Mizar Tilt高分辨率光片（最近推出的Tilt-RM） 共振镜，显微镜将减少“条纹”的一个固有缺陷，在光片 显微镜这一修改将大大增强系统的成像能力， 特别是用于定量荧光测量。

Summary The parent grant (1R35GM126980) for this equipment request supplement is entitled “Understanding Cell Division.” This award covers the primary research focus of the Gorbsky laboratory, analyzing the mechanics of chromosome movement, the regulation of the mitotic cell cycle, and the characterization of factors and conditions that contribute to chromosome instability. Much of our research involves high resolution imaging of cultured cells expressing fluorescently tagged proteins and are manipulated with RNA interference or CRISPR gene knockout and mutation. The Mizar Tilt light sheet provides high resolution imaging with minimal phototoxicity. The approaches used in the lab to interrogate protein function include siRNA or knocking degron/GFP tags into endogenous genes. We are also using the “Trim-away” technique, through which a combination of antibody and Trim13 E3 ubiquitin ligase, microinjected into living cells, allows rapid degradation of endogenous proteins without genetic manipulations. The requested upgrade to our Mizar Tilt high resolution light sheet, the recently introduced Tilt-RM resonance mirror, microscope will reduce “striping” an inherent defect in light sheet microscopy. This modification will greatly enhance the imaging capabilities of the system, particularly for quantitative fluorescence measurements.