Dysregulated transcription is frequently involved in cancer development. Chromatin immunoprecipitation (ChIP) assay is the technique of choice for examining in vivo transcription factor-DNA interactions. The current ChIP technology is limited by the requirement of a large number of cells and the long assay time caused by extensive manual processing. These problems practically prevent its use on primary cells extracted from animals and patients. In this project, we will develop microfluidic ChIP assays for studies based on tiny amounts of primary samples from mice and humans. We will achieve a sensitivity of ~20-50 cells for ChIP-qPCR and ~1000 cells for ChIP-seq for transcription factor binding studies using primary cells by the end of this project. Taking advantage of the ultrahigh sensitivity, we will demonstrate two innovative applications of ChIP assays: 1. Study transcriptional regulation in a cellular subset from primary tumors, namely tumor-initiating cells (TICs);2. Monitor temporal dynamics in transcriptional regulation by minimally-invasive examination of a single live mouse. These experiments cannot be conducted using current technology and will yield unique insights that improve the understanding of cancer development at the molecular level.

Public Health Relevance

Our goal is to develop a new generation of microfluidic chromatin immunoprecipitation assays that can be used for analysis of transcriptional regulation involved in cancer development using cell samples from animals and patients. The technology will generate unique insights into the molecular mechanisms involved in tumorigenesis and cancer development.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21CA174577-01
Application #
8471855
Study Section
Special Emphasis Panel (ZCA1-SRLB-Q (J1))
Program Officer
Li, Jerry
Project Start
2013-05-01
Project End
2016-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
1
Fiscal Year
2013
Total Cost
$256,488
Indirect Cost
$81,793
Name
Virginia Polytechnic Institute and State University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
003137015
City
Blacksburg
State
VA
Country
United States
Zip Code
24061
Ma, Sai; Loufakis, Despina Nelie; Cao, Zhenning et al. (2014) Diffusion-based microfluidic PCR for "one-pot" analysis of cells. Lab Chip 14:2905-9
Geng, Tao; Lu, Chang (2013) Microfluidic electroporation for cellular analysis and delivery. Lab Chip 13:3803-21