Microfluidic PCR System for Single Cell Transcriptional Analysis

Keller, Evan

Abstract

The objective of this proposal is to acquire a PCR system that is sensitive enough for single cell transcriptional analysis. The system, called the Biomark HD System from Fluidigm is capable of real- time, end-point and digital PCR. The key aspects of this system are the use of integrated microfluidic channels on one chip that allow for simultaneous PCR reactions in nanoliter volumes and picogram amounts of RNA. In concrete terms, transcripts (up to 96 per run) from individual cells (up to 96 per run) can be assayed in a single run. This unique capability will greatly expand the research questions that can be addressed at University of Michigan for multiple investigative directions. Specific examples include metastasis studies such as disseminated tumor cell biology or cancer stem cell biology. This application supports 14 major users who together have 20 R01s, 4 projects in one P01, 3 projects in two different P50 SPORES, 2 projects in U01 consortia and 1 each of a NIH Director's Award and a U54 award for a total of 31 NIH-funded projects. The group consists of members of multiple Departments and the Medical and Dental Schools. Currently, there is not capability to perform single cell transcriptional analysis at University of Michigan and this equipment will enable us to expand studies into this very important area. The equipment will be placed in a central location in the Cancer Center Building so that it is accessible to many investigators. The projects already planning to use this equipment will utilize 86.8% of the available equipment time indicating the high demand for this equipment. Research projects are directly related to NIH goals of improving health and include studies on cancer metastasis, cancer therapeutics, angiogenesis, bone biology and age- related disease. The equipment will be supported by an experienced technical staff, a bioinformatics expert and an internal advisory committee. A system to attract and train new users is in place so that other NIH-funded investigators will benefit from the equipment. Significant institutional support for both maintenance and technical costs has been committed for the initial three years to ensure long- term viability of the system (Total of $100,500 over 3 years). A user-based recharge system will be put into place to continue its long-term use. Additionally, there is institutional commitment of space for the equipment. In summary access to this cutting edge imaging system will greatly accelerate the pace of research at University of Michigan for a large number of NIH-funded investigators.