Abstract
The structure of chromatin in eukaryotes exerts significant influences on many DNA related processes, including transcription, replication, recombination and repair. A useful tool for mapping chromatin structure is micrococcal nuclease (MNase), which induces double-strand breaks within nucleosome linker regions, and with more extensive digestion, single-strand nicks within the nucleosome itself. Many studies, carried out largely with microbes and cell cultures, have used MNase to determine the positions of nucleosomes within a region of DNA to identify dynamic changes induced during gene regulation. To measure similar processes in a developmental context, we turned to a tractable model system, the Drosophila embryo. Here we describe a protocol that enables MNase mapping of the enhancer chromatin structure in the embryo, and show how it can be used to identify structural changes on a cis-regulatory element targeted by the Knirps repressor.
