Nucleic acid-binding properties of human low density lipoprotein

人低密度脂蛋白的核酸结合特性

基本信息

批准号：
7477782
负责人：
Natalia V. Guevara
金额：
$ 12.08万
依托单位：
UNIV/TEXAS BROWNSVILLE & SOUTHMOST COLL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7477782
关键词：
Affinity American American Type Culture Collection Avidity Binding Biological Assay Biotechnology Cell Line Cells Chinese Hamster Chinese Hamster Ovary Cell Clinical Protocols Collection Complex Cytomegalovirus DNA Data Digestion Electrophoretic Mobility Shift Assay Endothelial Cells Ensure Enzyme-Linked Immunosorbent Assay Epithelial Cells Family Fibroblasts Future Gene Delivery Genes Genetic Genome Goals Green Fluorescent Proteins Hela Cells Hepatitis C Hispanics Human Institution Knowledge Label Lipoprotein Binding Low-Density Lipoproteins Lung Mediating Neuroblastoma Nucleic Acid Binding Nucleic Acids Numbers Oligonucleotides Outcome Ovary Pilot Projects Plasmids Property Prostate Publications Publishing Purpose Range Replacement Therapy Reporter Genes Research Science Series Simplexvirus Standards of Weights and Measures Students Supervision Surveys Techniques Testing Time Tissues Training Transfection Vaccination Vaccines Viral Viral Genome Virus base cell type design design and construction gene delivery system gene replacement gene therapy human disease in vivo lipid transport particle promoter protein expression research study symposium tissue culture

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to explore and further develop a native low density lipoprotein (LDL)-based gene delivery system (AraVecTM) for the introduction of a broad range genetic constructs into various cell types. In these pilot studies we will perform a survey of cell types for which LDL can be used as a gene delivery vehicle. A number of cell lines, including HeLa cells, human fibroblasts, human MCF7s, human HEP1, human HepG2 cells, human neuroblastomas, human lung and endothelial cells, human prostate epithelial cells (LnCap), and Chinese hamster ovary CHO cell, will be obtained from American Type Culture Collection (ATCC). Commercially available plasmids containing green fluorescent protein (GFP) gene driven by human cytomegalovirus immediate early 2 (HCMV IE2) promoter will be used in these studies as a complex with native human LDL. Time course of LDL-DNA incorporation into cells and GFP expression will be determined for each cell type. We will also identify fragments of HCMV and Hepatitis C viral genomes that bind to LDL with high affinity. This project is a necessary step in developing experimental and clinical protocols for successful gene replacement therapies and DNA vaccines for a variety of debilitating or lethal human diseases. The data will be used in future studies where genetic constructs are designed with increased binding affinity to LDL and appropriate cell-specific expression. The data will also help to clarify in vivo functions of LDL other than lipid transport. The project is expected to increase research-based training capacities of the institution, and is aimed at engaging Hispanic students in cutting-edge biotechnology research. Graduate students and science majors will have an opportunity to participate in experimental studies under the PI's supervision, present their projects at national conferences, and co-author publications.

描述（由申请人提供）：该项目的目的是探索并进一步开发基于基于的天然低密度脂蛋白（LDL）基因输送系统（ARAVECTM），以将广泛的遗传构建体引入各种细胞类型。在这些试点研究中，我们将对可以将LDL用作基因输送工具的细胞类型进行调查。许多细胞系，包括HeLa细胞，人成纤维细胞，人MCF7，人HEP1，人HEPG2细胞，人神经母细胞，人肺和内皮细胞，人类前列腺上皮细胞（LNCAP）和中国仓鼠卵巢卵巢CHO，将从美国类型的养老院中获得。含有绿色荧光蛋白（GFP）基因的市售质粒，由人类巨细胞病毒驱动的基因立即2（HCMV IE2）启动子在这些研究中将用作与天然人LDL的复合物。 LDL-DNA掺入细胞的时间过程，将确定每种细胞类型的GFP表达。我们还将确定与高亲和力结合LDL的HCMV和丙型肝炎病毒基因组的片段。该项目是开发用于成功的基因替代疗法和DNA疫苗的实验和临床方案的必要步骤，用于各种衰弱或致命的人类疾病。该数据将在未来的研究中使用，其中设计遗传构建体具有增加与LDL的结合亲和力和适当的细胞特异性表达。数据还将有助于阐明除脂质传输以外的LDL的体内功能。该项目有望提高该机构的基于研究的培训能力，并旨在让西班牙裔学生参与尖端的生物技术研究。研究生和科学专业的专业将有机会在PI的监督下参加实验研究，在国家会议上介绍他们的项目以及合着者出版物。