Live Cell Correlative Imaging Probes

描述（由申请人提供）：将根据1.4 nm纳米金簇标签制备探针，用于活细胞的相关荧光和电子显微镜，这些探针可以在转染时不需要对细胞膜进行物理或化学破坏。将研究三种方法：通过结合渗透化学基团（如乙酰氧基甲基）或结合与蛋白质转导结构域（如HIV-1 Tat肽）相对应的肽，制备荧光和纳米金抗体联合探针的膜渗透类似物，以促进通过细胞膜的传递；以融合多组氨酸标签为靶点的抗体或硝基三乙酸基制备针对绿色和其他彩色荧光蛋白的膜透性金簇探针以及纳米金与FlAsH或其相应的结合肽的偶联物，分别针对所表达的肽或FlAsH标记的靶标。这些探针将用于哺乳动物细胞核中基因、RNA和表达蛋白的相关定位，以及细胞核中大规模染色质结构的研究。

DESCRIPTION (provided by applicant):Probes will be prepared, based on the 1.4 nm Nanogold cluster label, for the correlative fluorescence and electron microscopy of living cells, which may be transfected without requiring physical or chemical disruption of the cell membrane for transfection. Three approaches will be investigated: preparation of membrane permeable analogs of combined fluorescent and Nanogold antibody probes either by the incorporation of permeabilizing chemical groups such as acetoxymethyl, or by the conjugation of peptides corresponding to protein transduction domains such as the HIV-1 Tat peptide to facilitate delivery through cell membranes; preparation of membrane permeable gold cluster probes targeted to green and other colored fluorescent proteins by means of antibodies or nitrilotriacetic acid groups which target fused polyhistidine tags; and conjugates of Nanogold with FlAsH or with its corresponding binding peptide targeted respectively to the expressed peptide or to FlAsH-labeled targets. The new probes will be evaluated in the correlative localization of genes, RNA and expressed proteins in the mammalian cell nucleus, and to study the large-scale chromatin structure in the nucleus.