In vivo reporters of gene expression

基因表达的体内报告基因

基本信息

批准号：
8892080
负责人：
Marit Nilsen-Hamilton
金额：
$ 20.23万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-15 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8892080
关键词：
Actins Acute-Phase Proteins Acute-Phase Reaction Adverse effects Aminoglycosides Binding Biopsy Blood Blood Proteins Cells Cellular biology Chemicals Detection Deterioration Failure Gene Expression Gene Expression Profile Genes Graft Rejection Graft Survival Health Image Immune Immunosuppression Immunosuppressive Agents Incidence Inflammation Inflammatory Inflammatory Response Invaded Kidney Transplantation Life Ligands Location Maintenance Mammalian Cell Measures Molecular Biology Monitor Neomycin Organ Transplantation Organ failure Organic Chemistry Pharmaceutical Preparations Probability Procedures Radiochemistry Radiolabeled Reporter Reporter Genes Research Signal Transduction Staging Stem cells Stimulus Survival Rate System T-Lymphocyte Testing Tissues Transplantation Transplanted tissue Variant Veterans aptamer base cellular imaging imaging system in vivo macrophage novel strategies prevent promoter radioligand radiotracer receptor receptor binding response success trafficking

项目摘要

DESCRIPTION (provided by applicant): Organ transplants are becoming increasingly common and the one year graft survival rates have risen sharply over the past 20 years due to aggressive immunosuppressive drug treatments. For example, renal transplants now have one year success rates of over 90% in most transplant centers. However, the long-term survival rate has not increased significantly over this same period. Subclinical inflammation (SCI) is a major contributor to transplant deterioration and to increase long-term success rates of organ transplants it is vital to have a measure of SCI in the transplant tissue. SCI is currently assesse by the invasive measure of a biopsy but could be more effectively evaluated by imaging of gene(s) that are activated during inflammation. We have developed a gene expression reporter system that can be used for imaging gene expression in deep tissue. The reporters are aptamers that are expressed from a chosen promoter and are visualized when they bind their target radioligand. The aptamers behave as intracellular receptors, binding their ligands tightly and thereby providing an image of the cells in which the promoter has been activated. The current veteran team, which includes expertise in cell and molecular biology, synthetic organic chemistry and radiochemistry, will develop an aptamer reporter system for imaging local inflammation of transplanted tissues by achieving the following specific aims: 1) Prepare mammalian cells that stably express neomycin aptamers as reporters of either a regulated (Lcn2) or constitutively expressed (actin) gene, 2) Develop the chemical procedures to synthesize and radiolabel aptamer ligands, and 3) Image actin and Lcn2 gene expression in vivo. The studies proposed here have broad implications for monitoring the health of tissue transplants to enable more informed decisions regarding medications. Such monitoring is anticipated to increase the survival of transplants that have been modified by the inclusion of the aptamer SCI reporter system prior to their transplantation. The same reporter platform may be applied to other imaging needs such as stem cell trafficking or their gene expression profile to monitor their differentiation stage.

描述（由申请人提供）：器官移植变得越来越普遍，由于积极的免疫抑制药物治疗，在过去20年中，一年的移植物存活率急剧上升。例如，肾移植物现在在大多数移植中心的成功率超过90％。但是，在同一时期，长期存活率尚未显着提高。亚临床炎症（SCI）是导致移植变质的主要原因，并提高器官移植的长期成功率，在移植组织中拥有SCI至关重要。 SCI目前通过活检的侵入性度量进行评估，但可以通过在炎症期间被激活的基因成像进行更有效的评估。我们已经开发了一个基因表达报告基因，该系统可用于成像深组织中的基因表达。记者是从选择的启动子表达的适体，并在结合目标放射线时可视化。适体的表现为细胞内受体，紧密地结合其配体，从而提供了激活启动子的细胞图像。当前的资深团队包括细胞和分子生物学方面的专业知识，合成有机化学和放射化学，将开发一种适体报告基因系统，用于对移植组织的局部炎症进行成像，以实现以下特定目的：1）稳定地表达型neosycin Aptamer aptime a condement ote neyomycIn Aptame condement（或表达型号）。 2）开发化学程序以合成和放射性标记的适体配体，3）在体内图像肌动蛋白和LCN2基因表达。这里提出的研究对监测组织移植的健康具有广泛的影响，以实现有关药物的更明智的决定。预计这种监测将增加通过包含的移植物的存活 Aptamer SCI报告程序在移植之前。相同的记者平台可以应用于其他成像需求，例如干细胞运输或其基因表达谱以监测其分化阶段。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The Lipocalin2 Gene is Regulated in Mammary Epithelial Cells by NFκB and C/EBP In Response to Mycoplasma.

Lipocalin2 基因在乳腺上皮细胞中由 NFκB 和 C/EBP 调节以响应支原体。

DOI：
10.1038/s41598-020-63393-x
发表时间：
2020
期刊：
Scientific reports
影响因子：
4.6
作者：
Zhao,Wei;Bendickson,Lee;Nilsen-Hamilton,Marit
通讯作者：
Nilsen-Hamilton,Marit

Characterization of the Photophysical Behavior of DFHBI Derivatives: Fluorogenic Molecules that Illuminate the Spinach RNA Aptamer.

DOI：
10.1021/acs.jpcb.8b11166
发表时间：
2019-02
期刊：
The journal of physical chemistry. B
影响因子：
0
作者：
K. Santra;Ivan M Geraskin;M. Nilsen-Hamilton;G. Kraus;J. Petrich
通讯作者：
K. Santra;Ivan M Geraskin;M. Nilsen-Hamilton;G. Kraus;J. Petrich

Aptamer-enabled uptake of small molecule ligands.

DOI：
10.1038/s41598-018-33887-w
发表时间：
2018-10-24
期刊：
Scientific reports
影响因子：
4.6
作者：
Auwardt SL;Seo YJ;Ilgu M;Ray J;Feldges RR;Shubham S;Bendickson L;Levine HA;Nilsen-Hamilton M
通讯作者：
Nilsen-Hamilton M

Ligands with polyfluorophenyl moieties promote a local structural rearrangement in the Spinach2 and Broccoli aptamers that increases ligand affinities.