Systematic dissection and optimization of inducible enhancers in human cells using a massively parallel reporter assay.

ABSTRACT: Learning to read and write the transcriptional regulatory code is of central importance to progress in genetic analysis and engineering. Here we describe a massively parallel reporter assay (MPRA) that facilitates the systematic dissection of transcriptional regulatory elements. In MPRA, microarray-synthesized DNA regulatory elements and unique sequence tags are cloned into plasmids to generate a library of reporter constructs. These constructs are transfected into cells and tag expression is assayed by high-throughput sequencing. We apply MPRA to compare >27,000 variants of two inducible enhancers in human cells: a synthetic cAMP-regulated enhancer and the virus-inducible interferon-β enhancer. We first show that the resulting data define accurate maps of functional transcription factor b

SUBMITTER: Melnikov A 

PROVIDER: S-EPMC3297981 | biostudies-literature | 2012 Feb

REPOSITORIES: biostudies-literature