Genetically targeted fluorogenic macromolecules for subcellular imaging and cellular perturbation.
Genetically targeted fluorogenic macromolecules for subcellular imaging and cellular perturbation.
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DOI：
10.1016/j.biomaterials.2015.07.002
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发表时间：
2015-10
期刊：
影响因子：
14
通讯作者：
Bruchez MP
中科院分区：
文献类型：
--
作者：
Magenau AJ;Saurabh S;Andreko SK;Telmer CA;Schmidt BF;Waggoner AS;Bruchez MP
通过将大分子锚定到特定细胞器来改变细胞功能可能会揭示治疗潜力和临床治疗的新领域。在这项工作中，通过用遗传靶向大分子修饰亚细胞结构来影响细胞行为，从而诱发了独特的表型。这些荧光官能化聚合物通过受控自由基聚合制备，能够专门修饰表达局部荧光激活蛋白的活细胞的肌动蛋白、细胞质或核区室。通过建立影响结合率、结合亲和力和内化水平的关键聚合物结构-生物物理性质关系来优化大分子荧光剂。在不存在膜透化或转导域的情况下，在纳摩尔浓度的聚合物下实现了亚细胞结构的特异性标记，并且发现荧光修饰的聚合物在递送到细胞质后与完整的蛋白质结合。人们发现细胞运动依赖于大分子荧光剂与肌动蛋白结构的结合，导致细胞快速波动而不迁移。
The alteration of cellular functions by anchoring macromolecules to specified organelles may reveal a new area of therapeutic potential and clinical treatment. In this work, a unique phenotype was evoked by influencing cellular behavior through the modification of subcellular structures with genetically targetable macromolecules. These fluorogen-functionalized polymers, prepared via controlled radical polymerization, were capable of exclusively decorating actin, cytoplasmic, or nuclear compartments of living cells expressing localized fluorgen-activating proteins. The macromolecular fluorogens were optimized by establishing critical polymer architecture-biophysical property relationships which impacted binding rates, binding affinities, and the level of internalization. Specific labeling of subcellular structures was realized at nanomolar concentrations of polymer, in the absence of membrane permeabilization or transduction domains, and fluorogen-modified polymers were found to bind to protein intact after delivery to the cytosol. Cellular motility was found to be dependent on binding of macromolecular fluorogens to actin structures causing rapid cellular ruffling without migration.