Transcription Factor Reporter Technology

描述（由申请人提供）：癌症基因表达途径的分子解剖揭示了许多癌症治疗的新靶点。 这些目标包括异常转录机制的组成部分。 由于许多信号转导途径在转录水平上的会聚，参与转录调控的蛋白质获得高度优先。 针对转录靶点的新分子疗法由于其干扰靶基因表达的精确性而比传统疗法具有显著优势。 因此，正在开发小分子抑制剂、DNA结合聚酰胺、蛋白质结合寡核苷酸诱饵以及小干扰RNA及其组合用于癌症治疗。 虽然分子遗传学和药物化学的快速进展提供了新的基因表达“衰减剂”，但目前缺乏适合这些疗法的癌症早期和非侵入性评估技术。 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。 拟议的研究的目标是优化和表征远红荧光染料标记的寡脱氧核苷酸分子报告探针（ODMR），然后调查其转录因子的报告属性。 本论文以NF-kB作为肿瘤相关转录因子的模型，主要研究了以下几个方面：1）转录因子报告探针（ODMR）的设计、合成和体外检测; 2）ODMR探针检测活细胞中活性转录因子的潜力。

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.