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.
