GammaPNA Miniprobes for Telomere Analysis and RNA FISH

用于端粒分析和 RNA FISH 的 GammaPNA 微型探针

基本信息

批准号：
9047550
负责人：
Bruce A. ARMITAGE
金额：
$ 48.45万
依托单位：
PNA INNOVATIONS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9047550
关键词：
Address Affinity Age Archives Behavioral Research Biological Assay Biological Markers Biological Sciences Biology Biomedical Research Businesses Cell Line Cell Nucleus Cell Proliferation Cell division Cells Centromere Chemistry Chromosomal Stability Chromosomes Clinical Color Complementary DNA DNA DNA Damage Data Detection Development Diagnostic Diagnostics Research Disease Epidemiologic Studies Exhibits Feedback Fluorescence Fluorescence Resonance Energy Transfer Fluorescent Dyes Fluorescent Probes Fluorescent in Situ Hybridization Functional disorder Generations Goals Hand Health Human Inherited Interphase Jurkat Cells Label Laboratories Lead Left Length Libraries Longevity Malignant Neoplasms Marketing Messenger RNA Methods Modification Molecular Natural regeneration Paper Pathology Peptide Nucleic Acids Performance Phase Proteins Publications Publishing RNA RNA Sequences RNA analysis RNA library Research Research Personnel Resolution Sales Sampling Signal Transduction Site Solubility Staining method Stains Technology Telomerase Telomere Length Maintenance Testing Tissue Sample Tissues Transgenic Mice Universities Vertebral column Work age related analog base biophysical properties cancer risk design disorder risk epidemiology study follow-up human disease improved innovation interest member monomer public health relevance research study response success telomere tool

项目摘要

DESCRIPTION (provided by applicant): The proposed Phase II project will continue development of gammaPNA miniprobe technology originally developed at Carnegie Mellon University and the University of Pittsburgh and subsequently transferred to PNA Innovations, Inc, a small business spun out of Carnegie Mellon University. The basis of gammaPNA miniprobes is the high affinity with which gammaPNA hybridizes to complementary DNA. The specific applications addressed in this proposal are telomere analysis, which is currently done using fluorescent PNA probes 18 bases in length, which hybridize to 3 consecutive repeats of the human telomere sequence 5'-AATGGG-3', and mRNA labeling by complementary fluorescent probes. In Phase I, we demonstrated that the higher affinity of gammaPNA allows shorter 12 base telomere probes to be used, resulting in more fluorescent dyes being delivered to a telomere of a given length. This allow more reliable analysis of the shortest (i.e. critically short) telomeres, which are implicated in a variety of conditions including aging-related diseases and cancer. We published a paper describing our results in Organic and Biomolecular Chemistry and we launched a marketing campaign around our Telo MiniprobesTM, leading to our first sales within this product line. The proposed research will have four Specific Aims. The first aim extends our Phase I work in three ways. First we will study several additional cell lines This will help us to (a) determine the range of variability in miniprobe performance and (b) potentially identify other versions of the miniprobe that work in varied cell lines. Second, we wil develop miniprobes that target the C-rich telomere strand. Third, we will build on promising preliminary results for synthesizing internally labeled miniprobes that will double or triple the brightness of our current best probe. The second and third aims are directed toward new applications, specifically in development of a high throughput telomere assay based on our miniprobes and testing of fresh and archived tissue samples, which are currently difficult to study by FISH due to autofluorescence. The fourth aim will significantly extend gammaPNA FISH probes into RNA labeling. We will synthesize sets of gammaPNAs targeted to different sites on a single mRNA, but rather than covalently label the probes, which is costly, we will use an innovative and economical co-hybridization approach to label our probes. The synthesis of the gammaPNA monomers and oligomers will be done at PNA Innovations. Biophysical characterization and telomere staining will be done at academic laboratories at Carnegie Mellon and the University of Pittsburgh where the gammaPNA miniprobe technology was invented. Optimized miniprobes will then be sent to independent beta-testing laboratories that currently use conventional PNA probes for telomere analysis or DNA-based molecular beacons for mRNA labeling.

描述（由申请人提供）：gampna Miniprobe技术的继续开发，最初开发了Carnegie Mellon和Pittsburgh大学。 γ杂交杂交与同级DNA。在OPOSAL中的特定应用地址是端粒分析，目前使用荧光PNA探针长度为18个碱基，该碱基的长度为3'Uman端粒序列5'-TGGG-3'WS Shorters Shorters Shorter使用，导致更多的I荧光染料被传递到给定长度的端粒。简称）与衰老相关的疾病和癌症有关，我们围绕电视迷你菜单发起了一场营销活动，从而导致我们在Thin The toiss产品线中首次销售。三种方式。合成的亮度或三重探针的内部标记的微型生物。由于自动荧光而导致的第四个目标会显着将γ鱼探针扩展到单个mRNA上的RNA标记。标记我们的探针。用于mRNA标记的基于DNA的分子信标。