UNS: Engineering protein sensors to detect epigenetic changes during stem cell culture

UNS：工程蛋白质传感器检测干细胞培养过程中的表观遗传变化

• 批准号: 1512285
• 负责人: Chongli Yuan
• 金额: $ 30万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1512285&HistoricalAwards=false
• 关键词: UNS; Engineering; protein; sensors; detect

1512285 Yuan, Chongli DNA methylation is important in regulating gene activity , and in a cell population cells may have exhibit different levels of methylation, which results in behaviors that vary from the population average. Understanding the methylation level changes in individual cells is important in many biological applications. In this project the researchers will develop a novel method which will enable measurement of methylation at a single-cell level. Intellectual Merit: Current detection approaches for quantifying DNA methylation are limited because they preclude single-cell analysis and real-time monitoring of epigenetic changes. The researchers propose to design a probe by combining protein engineering principles and modern fluorescence detection techniques. Three objectives will be pursued: engineer and validate molecular sensors that detect methylated DNA near a gene locus, optimize the probes for use with cell extracts, and evaluate the genetically-encoded probe within live cells. Additionally, methylation level changes during stem cell passaging will also be measured. Broader Impacts: The proposed probe has the capability to significantly improve detecting DNA methylation levels at a single cell and potentially enable monitoring of dynamic changes. The research project provides a multidisciplinary training opportunity for engineers. The PIs will involve undergraduate students in research and encourage the participation of minority and female students. This proposal will also support the continued collaboration with the Women in Engineering Program and develop a new laboratory module for engaging students with the concept of fluorescence-based detection techniques.

1512285元，崇礼DNA甲基化是调节基因活性的重要因素，在一个细胞群体中，细胞可能表现出不同程度的甲基化，这导致行为与群体平均水平不同。了解单个细胞中甲基化水平的变化在许多生物学应用中都很重要。在这个项目中，研究人员将开发一种新的方法，使在单细胞水平上测量甲基化成为可能。智力优势：目前量化DNA甲基化的检测方法是有限的，因为它们排除了对表观遗传变化的单细胞分析和实时监测。研究人员建议将蛋白质工程原理和现代荧光检测技术结合起来，设计一种探针。将追求三个目标：设计和验证检测基因位点附近甲基化DNA的分子传感器，优化用于细胞提取物的探针，以及评估活细胞内的遗传编码探针。此外，还将测量干细胞传代过程中甲基化水平的变化。更广泛的影响：建议的探针能够显著提高对单个细胞DNA甲基化水平的检测，并可能实现对动态变化的监测。该研究项目为工程师提供了一个多学科的培训机会。PIs将让本科生参与研究，并鼓励少数族裔和女性学生参与。这项提案还将支持继续与妇女参与工程方案合作，并开发一个新的实验室模块，让学生了解基于荧光的检测技术的概念。

项目成果

Pre-differentiation exposure to low-dose of atrazine results in persistent phenotypic changes in human neuronal cell lines

分化前暴露于低剂量莠去津会导致人类神经细胞系持续表型变化

• DOI: 10.1016/j.envpol.2020.116379
• 发表时间: 2021
• 期刊: Environmental Pollution
• 影响因子: 8.9
• 作者: Xie, Junkai;Lin, Li;Sánchez, Oscar F.;Bryan, Chris;Freeman, Jennifer L.;Yuan, Chongli
• 通讯作者: Yuan, Chongli

Review: Engineering in situ biosensors for tracking cellular events

• DOI: 10.1016/j.coche.2020.06.006
• 发表时间: 2020-12
• 期刊: Current opinion in chemical engineering
• 影响因子: 6.6
• 作者: Kyle Wettschurack;Junkai Xie;Oscar F. Sánchez;Chongli Yuan

Identifying distinct heterochromatin regions using combinatorial epigenetic probes in live cells

• DOI: 10.1016/j.bbagrm.2021.194725
• 发表时间: 2021-06-26
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS
• 影响因子: 4.7
• 作者: Mendonca，Agnes;Sanchez，Oscar F.;Yuan，Chongli
• 通讯作者: Yuan，Chongli