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.
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.
|Sun, Chen; Hsieh, Yuan-Pang; Ma, Sai et al. (2017) Immunomagnetic separation of tumor initiating cells by screening two surface markers. Sci Rep 7:40632|
|Cao, Zhenning; Lu, Chang (2016) A Microfluidic Device with Integrated Sonication and Immunoprecipitation for Sensitive Epigenetic Assays. Anal Chem 88:1965-72|
|Sun, Chen; Hassanisaber, Hamid; Yu, Richard et al. (2016) Paramagnetic Structures within a Microfluidic Channel for Enhanced Immunomagnetic Isolation and Surface Patterning of Cells. Sci Rep 6:29407|
|Ma, Sai; Bryson, Bryan D; Sun, Chen et al. (2016) RNA Extraction from a Mycobacterium under Ultrahigh Electric Field Intensity in a Microfluidic Device. Anal Chem 88:5053-7|
|Cao, Zhenning; Chen, Changya; He, Bing et al. (2015) A microfluidic device for epigenomic profiling using 100 cells. Nat Methods 12:959-62|
|Loufakis, Despina Nelie; Cao, Zhenning; Ma, Sai et al. (2014) Focusing of mammalian cells under an ultrahigh pH gradient created by unidirectional electropulsation in a confined microchamber†Electronic supplementary information (ESI) available: Figures S1-S5 and videos S1-S2. See DOI: 10.1039/c4sc00319eClick here fo Chem Sci 5:3331-3337|
|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|