Skip to main content
. Author manuscript; available in PMC: 2013 Nov 14.
Published in final edited form as: Methods Mol Biol. 2013;1042:10.1007/978-1-62703-526-2_16. doi: 10.1007/978-1-62703-526-2_16

Fig. 1.

Fig. 1

System for monitoring communication between DNA damage responses and local transcription. (a) Schematic of reporter. The mCherry-LacI-Fok1 fusion protein creates DSBs within 256 lac operator repeats. Transcription is inducible upon doxycycline (Dox) addition, which activates a CFP gene. YFP-MS2 focally accumulates at the locus by binding to stem–loop structures within the 3′-UTR of the nascent transcript. (b) Expression of wild-type mCherry-LacI-Fok1 (FokI WT), but not nuclease-inactive FokI D450A, induces DSBs as indicated by gH2AX focus formation. (c) YFP-MS2 accumulates at the locus in FokI D450A-expressing cells indicating the presence of nascent transcription. This does not occur in FokI WT cells indicating that DSB responses silence transcription