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. 2012 Aug 10;5(6):956–966. doi: 10.1242/dmm.009696

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© 2012. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms.

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Fig. 1. — Rosa26 promoter analysis and design of deletions for signaling sentinel substitution. (A) Scheme depicts the Rosa26 locus; arrow, transcribed region. Boxes above depict sequence identities over 95% and 80% for mouse and rat (mo:ra) and mouse and human (mo:hu), respectively. Red bars within the boxes indicate conserved domains, largely extending to −3 kb of the start site of transcription. (B) VISTA plot of transcription factor motif distribution across the proximal 3 kb of the mouse Rosa26 promoter region. Vertical dotted line indicates a marked increase in motif density at −1.2 kb. (C) Comparison of the mouse and human Rosa26 promoter regions. A natural break occurs at −228 bp and a CCAAT box is evident downstream of −60 bp. (D) RMCE at the Rosa26 locus (upper scheme) results in substitutions that replace the regions from −4 kb to +1 kb with recombinant sequences where signaling response sentinels (green boxes) are inserted between deletion endpoints −60, −228, −1217 and −3000 and a TATA box fused to a coding sequence for CFP.