CRISPR Microfluidic Array for Simultaneous Multiplexed Sensing of Proteins and miRNAs

用于同时多重检测蛋白质和 miRNA 的 CRISPR 微流体阵列

• 批准号: 2231490
• 负责人: James Rusling
• 金额: $ 49.96万
• 依托单位: University of Connecticut
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2231490&HistoricalAwards=false
• 关键词: CRISPR; Microfluidic; Array; Simultaneous; Multiplexed

Measurement of biomolecules like proteins and nucleic acids, called biomarkers, in blood, is a valuable approach to detect cancers and other diseases. Samples consisting of several proteins and nucleic acids show promise for this purpose, but at present must be measured using separate, time-consuming methods. This project uses a biological construct called CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) to enable measurements. CRISPR can be designed to bind a nucleic acid or a protein labeled with a nucleic acid and give a signal only when that specific nucleic acid or label binds to CRISPR. This approach will be developed into a fast, low-cost 3D-printed, semi-automated device to measure several nucleic acids and several protein biomarkers in the same test. The project will also develop sensitive electrically-driven light emission called electrochemiluminescence (ECL) as a simpler detection method. The blood test using the 3D-printed device will be designed to run in a short time in a clinic or physician’s office by a lightly trained technician. This will allow fast turnaround in what is called point of care mode. Results will be available to the physician while the patient is still in his or her office and can be used at that time as part of the diagnosis. This project will utilize programs at Univ. of Connecticut to support education in new CRISPR technology for graduate students, undergraduates, science teachers and high school students. Activities include holding workshops on CRISPR detection through SECRET (School of Exploratory Chemistry Research Experience and Training), developing Early College Experience programs for secondary school students and science teachers, facilitating training of Ph.D. students in CRISPR array design and fabrication, and providing research opportunities for undergraduates at UCONN and in UCONN’s NSF REU program, as well as high school students through the ACS SEED program.Molecular biomarkers have become increasingly valuable in medical diagnostics for many diseases. Panels of biomarkers, as opposed to single biomarkers, show superior diagnostic utility and combined protein-miRNA panels have shown significantly improved diagnostic predictions over separate protein or miRNA panels. This project will develop the first analytical array using CRISPR technology to simultaneously measure combinations of miRNAs and proteins. The new 3D-printed array will be able to analyze miRNA-protein biomarker panels for a wide range of disease diagnostics in point-of-care (POC) mode, as well as in other biomedical research applications. In addition to fluorescent detection, electrochemiluminescence (ECL) will be explored with CDC and iPhone camera detection. The focus will be on miRNA and protein biomarkers that are high performing biomarkers for aggressive prostate cancer. Assays will be validated by measuring target miRNAs and proteins in 130 existing human serum samples from patients with high grade prostate cancers and controls. The intellectual merit of this project involves design and development of the first engineering strategy using a new CRISPR assay technology to detect panels of miRNAs and protein simultaneously at POC.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

测量蛋白质和核酸（称为生物标志物）的生物分子是一种检测癌症和其他疾病的宝贵方法。由几种蛋白质和核酸组成的样品对此目的显示出希望，但必须使用单独的耗时方法来测量。该项目使用称为CRISPR的生物学结构（群集定期间隔短的短粒子重复序列）来实现测量。 CRISPR可以设计用于结合核酸或用核酸标记的蛋白质，并且只有在该特定的核酸或标签与CRISPR结合时才能发出信号。在同一测试中，这种方法将发展为快速，低成本的3D打印，半自动化装置，以测量几种核酸和几种蛋白质生物标志物。该项目还将开发出敏感的电气发射发射，称为电化学发光（ECL）作为一种更简单的检测方法。使用3D打印设备的血液测试将设计为训练有素的技术人员在诊所或物理办公室的短时间内进行。这将允许在所谓的护理点模式下快速周转。当患者仍在他或她的办公室时，结果将可供物理，当时可以用作诊断的一部分。该项目将在大学使用计划。康涅狄格州为研究生，本科生，科学老师和高中生提供新的CRISPR技术教育。活动包括通过秘密（探索化学研究经验和培训学院）举办有关CRISPR检测的研讨会，为中学生和科学老师开发早期的大学经验计划，支持博士学位培训。 CRISPR阵列设计和制造的学生，为UConn和UConn的NSF REU计划提供研究机会，并通过ACS Seed Program计划为高中生提供研究。各分子生物标志物在许多疾病的医学诊断中变得越来越有价值。与单个生物标志物相反，生物标志物的面板表现出优越的诊断效用和蛋白质miRNA面板的优越性，已经显示出与单独的蛋白质或miRNA面板相比，诊断预测显着改善了诊断预测。该项目将开发使用CRISPR技术的第一个分析阵列，以同时测量miRNA和蛋白质的组合。新的3D打印阵列将能够分析miRNA-蛋白质生物标志物面板，以在护理点（POC）模式以及其他生物医学研究应用中进行广泛的疾病诊断。除荧光检测外，还将通过CDC和iPhone相机检测探索电化学发光（ECL）。重点将放在具有侵略性前列腺癌的高性生物标志物的miRNA和蛋白质生物标志物上。测量将通过测量来自高级前列腺癌和对照患者的130例现有人血清样品中的靶标miRNA和蛋白质来验证。该项目的智力优点涉及使用新的CRISPR分析技术设计和开发第一个工程策略的设计和制定，以在POC上仅在POC上检测MiRNA和蛋白质面板。该奖项反映了NSF的法定任务，并通过使用基金会的知识分子优点和广泛的审查标准来评估，通过评估来诚实地支持。