Transcription Factor Reporter Technology

描述(申请人提供)：对癌症基因表达途径的分子解剖揭示了癌症治疗的许多新靶点。这些靶点包括异常转录机器的组件。由于许多信号转导途径在转录水平上的汇聚，参与转录调控的蛋白质得到了高度重视。与传统疗法相比，针对转录靶点的新分子疗法具有显著的优势，因为它们可以精确地干扰靶基因的表达。因此，小分子抑制剂、DNA结合聚酰胺、蛋白质结合寡核苷酸诱饵以及小干扰RNA及其组合正在被开发用于癌症治疗。虽然分子遗传学和药物化学的快速发展为基因表达提供了新的“衰减器”，但目前还缺乏对癌症进行早期和非侵入性评估的技术，这些技术适用于这些疗法。特别是，直接报告癌细胞中基因转录的成像技术对于癌症表型和分期以及评估新疗法都至关重要。本研究的目的是优化和表征远红光荧光标记的寡核苷酸分子报告探针(ODMR)，并研究其转录因子报告特性。利用核因子-kB作为癌症相关转录因子的模型，将追求以下特定目标：1)转录因子报告探针(ODMR)的设计、合成和体外测试的优化；2)研究ODMR探针在检测活细胞中活性转录因子方面的潜力。

DESCRIPTION (provided by applicant): Molecular dissection of gene expression pathways in cancer has revealed many new targets for cancer therapy. Those targets include the components of abnormal transcription machinery. Proteins involved in regulation of transcription attained high priority due to the convergence of many signal transduction pathways at the transcriptional level. New molecular therapies directed to transcriptional targets have significant advantages over traditional therapies due to a precision of their interference with target gene expression. Consequently, small molecule inhibitors, DNA binding polyamides, protein-binding oligonucleotide decoys, as well as small interfering RNAs and their combinations are being developed for cancer therapy. While rapid progress in molecular genetics and medicinal chemistry delivers new "attenuators" of gene expression, the technologies of early and non-invasive assessment of cancers that would be amenable to these therapies are currently lacking. In particular, imaging technologies that report directly on gene transcription in cancer cells are critically important for both cancer phenotyping and staging, as well as for evaluating new therapies. The goal of the proposed research is to optimize and characterize far-red fluorochrome labeled oligodeoxyribonucleotide molecular reporter probes (ODMRs) followed by the investigation of their transcription-factor reporting properties. Using NF-kB as a model cancer-relevant transcription factor, the following specific aims will be pursued: 1) Optimization of design, synthesis, and in vitro testing of transcriptional factor reporter probes (ODMR); 2) Investigation of the potential of ODMR probes in detecting active transcription factor in live cells.