Baculovirus-encoded protein expression for epigenomic profiling in Drosophila cells.
The expression and genome-wide mapping of epitope-tagged DNA- and chromatin-binding proteins in cultured cells has become a powerful strategy for epigenome characterization, especially in Drosophila, where cell lines derived from numerous tissues are now available. However this strategy relies on establishing transfected cell lines, which is time-consuming and introduces variability. Here we show that baculovirus-encoded proteins can be efficiently produced following infection of Drosophila cell lines of different types. Using chromatin affinity purification, we show that epitope-tagged proteins produced in baculovirus-infected cells provide genome-wide profiles of the histone variant H2Av that are comparable to those produced by plasmid-transfected cells. The ability to express multiple epitope-tagged proteins for epigenome analysis from a single culture, and to do this in a variety of Drosophila cell lines, significantly extends the range of epigenome analysis.
Bryson
Terri D
TD
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Weber
Christopher M
CM
Henikoff
Steven
S
D000818:Animals; D016367:Baculoviridae / Q000502:physiology*; D019921:Biotinylation; D002460:Cell Line; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D057890:Epigenomics / Q000379:methods*; D006657:Histones / Q000378:metabolism; D010957:Plasmids; D014176:Protein Biosynthesis
D006657:Histones