Pre-mRNA Trans-Splicing for Molecular Imaging of Cancer

用于癌症分子成像的前 mRNA 反式剪接

• 批准号: 6603797
• 负责人: GERARD J. MC GARRITY
• 金额: $ 21.86万
• 依托单位: INTRONN, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6603797
• 关键词: RNA splicing; bioimaging /biomedical imaging; chorionic gonadotropin; diagnosis design /evaluation; epidermal growth factor; genetic markers; growth factor receptors; human papillomavirus; imaging /visualization /scanning; luciferin monooxygenase; metastasis; molecular cloning; neoplasm /cancer diagnosis; neoplasm /cancer genetics; oncogenes; spliceosomes; technology /technique development

New technologies are needed to facilitate the analysis, quantitation and imaging of gene expression to exploit the wealth of information being generated, for example, by the Human Genome and Cancer Genome Anatomy Procect. The innovative use of Spliceosome Mediated RNA Trans-splicing (SmaRT) to develop new agents for real time imaging of gene expression within cells, animals, and potentially humans is proposed. Our laboratory has produced different RNAs, known as PTMs (Pre-Trans-splicing Molecules). These PTMs are capable of directing trans-splicing reactions between the PTM and a targeted pre-messenger RNA. The product of a SmaRT reaction is a novel chimeric mRNA, which can encode virtually any desired gene product that may be imaged directly or that can activated or capture a second reporter molecule. The product of a SMaRT reaction contains one or more exons of the target endogenous pre-mRNA and a exonic or cDNA sequence delivered by the PTM. We propose studies to target cancer specific or cancer associated genes: human papillomavirus, human chorionic gonadotropin and EGF receptor with PTMs encoding marker luciferase exons. This application proposes to define the collaboration with the imaging group at the UCLA Medical Center to provide guidance on more advanced imaging techniques and equipment. Our specific aims include the demonstration that SMART can target clinically relevant genes with the luciferase marker at the pre-mRNA level. Subsequent studies will increase the efficiency and specificity of the PTMs through a molecular library. Real time molecular imaging. Real time molecular imaging by spliceosome mediated RNA trans-splicing will greatly facilitate pre-clinical animal studies in cancer gene single copy PCR detection. It may also imaging of tissue distribution of transgenes and vectors, an attractive alternative to single copy PCR detection. It may also be possible to identify metastases in small animal cancer models. If successful these animal studies could lead to human diagnostics of real time cancer specific RNA profiles. SMaRT is attractive single it targets RNA and holds the potential for real time analysis of gene expression.

需要新的技术来促进基因表达的分析、定量和成像，以利用例如由人类基因组和癌症基因组解剖过程产生的丰富信息。提出了剪接体介导的RNA反式剪接（SmaRT）的创新用途，以开发用于细胞、动物和潜在人类内基因表达的真实的时间成像的新试剂。我们的实验室已经产生了不同的RNA，称为PTM（前反式剪接分子）。这些PTM能够指导PTM和靶向的前信使RNA之间的反式剪接反应。SmaRT反应的产物是新的嵌合mRNA，其可以编码几乎任何所需的基因产物，所述基因产物可以直接成像或可以活化或捕获第二报道分子。SMaRT反应的产物含有靶内源性前mRNA的一个或多个外显子和由PTM递送的外显子或cDNA序列。我们建议研究靶向癌症特异性或癌症相关基因：人乳头瘤病毒，人绒毛膜促性腺激素和EGF受体与PTM编码标记荧光素酶外显子。本申请旨在定义与UCLA医学中心成像组的合作，以提供更先进成像技术和设备的指导。我们的具体目标包括证明SMART可以在前mRNA水平用荧光素酶标记物靶向临床相关基因。后续研究将通过分子文库提高PTM的效率和特异性。真实的实时分子成像。利用剪接体介导的RNA反式剪接进行真实的实时分子成像，将极大地促进肿瘤基因单拷贝PCR检测的临床前动物研究。它也可以成像的转基因和载体的组织分布，一个有吸引力的替代单拷贝PCR检测。也可以在小动物癌症模型中识别转移。如果成功的话，这些动物研究可能导致人类诊断真实的癌症特异性RNA谱。SMaRT是一种有吸引力的靶向RNA的方法，具有真实的实时基因表达分析的潜力。