SINGLE PRIMER MEDIATED CLONING OF P53 REGULATED GENES

通过第一阶段SBIR的支持，我们成功地证明了 使用一种新颖和创新的专有技术， 鉴定肿瘤转录控制下的基因 抑制基因p53。这项技术有几个明显的优势， 目前用于回收DNA序列的方法，如文库 筛选和PCR。我们利用这项技术分离出两个基因 在前列腺、睾丸、结肠和卵巢上皮细胞中表达 组织. 该提案的直接目标是：1）开发体内 评价这些基因的p53-反应性的生物学测定和2） 以确定这些基因的异常表达是否与 不受控制的增殖和锚定独立生长。 我们假设，如果p53调节基因在或接近于 多个信号转导通路的汇合点，它们可能是 细胞增殖状态的高度特异性标志物。因此，在本发明中， 它们的不适当表达可能在发展中具有重要意义， 癌症的进展。这些基因的分离和鉴定 可能导致识别特定的诊断和预后 许多癌症的指标，他们可能有一个潜在的 开发新的治疗试剂。 拟议的商业应用：目前可用的癌症标志物不 提供足够的诊断和预后信息。最 这项研究的重要商业应用将是 开发高度特异性的致瘤标记物， 早期诊断和癌症患者的临床评估。

Through the support of a Phase I SBIR we have successfully demonstrated the use of a novel and innovative proprietary technology for the identification of genes under the transcriptional control of the tumor suppressor p53. This technology has several distinct advantages over current methods used in the recovery of DNA sequences, such as library screening and PCR. We have used this technology to isolate two genes which are expressed in prostate, testis, colon, and ovarian epithelial tissue. The immediate objectives of this proposal are 1) to develop an in vivo biological assay to evaluate the p53-responsiveness of these genes and 2) to determine if aberrant expression of these genes is associated with uncontrolled proliferation and anchorage independent growth. We hypothesize that if p53 regulated genes function at or near the convergence point of multiple signal transduction pathways, they may be highly specific markers of the proliferative status of a cell. Thus, their inappropriate expression may be significant in the development and progression of cancer. The isolation and characterization of these genes may lead to the identification of specific diagnostic and prognostic indicators for a number of cancers, and they may have a potential for the development of novel therapeutic reagents. PROPOSED COMMERCIAL APPLICATION: Current available cancer markers do not provide adequate diagnostic and prognostic information. The most important commercial application of this research would be the development of highly specific tumorigenic markers which could be used in early diagnosis, and in the clinical evaluation of cancer patients.