title: Microfluidics Technologies for Low Cell Number Chromatin Immunoprecipitation.
abstract: Protein-DNA interactions are responsible for numerous critical cellular events: For example, gene expression and silencing are mediated by transcription factor protein binding and histone protein modifications, and DNA replication and repair rely on site-specific protein binding. Chromatin immunoprecipitation (ChIP) is the only molecular assay that directly determines, in a living cell, the binding association between a protein of interest and specific genomic loci. It is an indispensible tool in the biologist's toolbox, but the many limitations of this technique prevent broad adoption of ChIP in biological studies. The typical ChIP assay can take up to 1 wk to complete, and the process is technically tricky, yet tedious. The ChIP assay yields are also low, thus requiring on the order of millions to billions of cells as starting material, which makes the assay unfeasible for studies using rare or precious samples. For example, fluorescence-activated cell sorting (FACS) of cancer stem cells (CSCs) obtained from primary tumors, rarely yields more than ~100,000 CSCs per tumor. This protocol describes a microfluidics-based strategy for performing ChIP, which uses automation and scalability to reduce both total and hands-on assay time, and improve throughput. It allows whole fixed cells as input, and enables automated ChIP from as few as 2000 cells.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, California 94305;
authors.forename: Angela R
authors.initials: AR
authors.lastname: Wu
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, California 94305; Howard Hughes Medical Institute, Stanford, California 94305.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Quake
mesh_terms: D047369:Chromatin Immunoprecipitation / Q000379:methods*; D044085:Microfluidics / Q000379:methods*