Project Summary Single-cell RNA sequencing (scRNA-seq) holds the promise to revolutionize the study of biology by allowing measurement of cellular processes within individual cells with unprecedented precision and breadth. Micro-RNAs (miRNA) are a class of small regulatory RNAs that are involved in a range of biological processes such as organismal development, cellular differentiation, and major signaling pathways. miRNA signatures correlate strongly with disease phenotype and can serve as reliable biomarkers. This proposal will develop a microfluidic platform that can allow isolation and sequencing of miRNA from individual single-cells in a highly-parallel and unbiased manner. In the K99 phase, I will sequentially develop each segment of the device and optimize the reactions and processes associated with it. The device will be tested and validated using control cell lines to determine the sensitivity of detection, and establish the quality of single cell data. Finally, I will use the platform to profile miRNA expressions in leukemia cell lines and those obtained from patients with the goal to identify expression patterns that will have diagnostic or prognostic value. The R00 phase of the project will focus on applying this platform in studying disease pathogeneses in leukemia. Findings from the study will shine light on new pathways for disease progression and drug resistance, which can be used for developing new therapeutic modalities. I will also develop the platform as an ultra-sensitive diagnostic tool for detecting leukemia in patients. I also propose an extensive training program that will help me realize my career goals and support my transition to an independent research position. The research environment at Koch Institute is unparalleled and offers the unique opportunity to interact and collaborate with eminent biologists and engineers. I have assembled an exceptional team: Prof. Dan Anderson and Prof. Robert Langer, experts in bioengineering and high-throughput techniques, will mentor me. I will work very closely with Prof. Phillip Sharp, RNA Biologist, who will help me address the molecular biology component of the project. Dr. Amir Faithi will provide clinical samples and direction to increase the medical impact of the project. My training will also involve mentoring students, presenting my work in single-cell meetings, and science outreach.