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. 2014 Feb 21;42(8):4906–4921. doi: 10.1093/nar/gku139

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© The Author(s) 2014. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — Transgenes designed to identify structural features at the hGH locus critical to placental gene activation. (A) The hGH locus and epigenetic modifications in the placenta. The hGH gene cluster encompasses five genes: the pituitary-specific growth hormone gene (hGH-N), and four placenta-specific paralogs, hCS-L, hCS-A, hGH-V and hCS-B. The gene cluster is flanked 5′ by SCN4A and CD79b genes and 3′ by TCAM1. These flanking genes are specifically expressed in skeletal muscle, B lymphocyte and testis, respectively. Our previous studies identified two overlapping sets of DNase I HSs located 5′ to the cluster in the pituitary and placenta. HSIII and V are present in multiple tissues, including pituitaries and placentas, HSI and II are pituitary-specific and critical to hGH-N expression, and HSIV is specific to the placenta (4). Previously established patterns of histone H3 and H4 acetylation and H3K4 di- and tri-methylation at the hGH locus in the chromatin of human placental STBs are indicated (5). (B) Three hGH/BAC-derived transgenes designed to identify structural determinants of placental gene expression. The structures of four transgenes are shown. The wild-type hGH/BAC transgene (hGH/BAC) comprises a 123-kb NotI genomic fragment containing the entire hGH cluster with extensive 5′- and 3′-flanking regions, including the full set of pituitary or placental LCR determinants. The HSIII–V region (placental LCR) was selectively deleted from the hGH/BAC to generate the ΔHSIII-V/hGH/BAC transgene. A 12-kb segment between HSIII and the hGH cluster was deleted from hGH/BAC to generate the ΔSpacer/hGH/BAC transgene. In transgene LCR-CSA/BAC, the entire hGH cluster was replaced by a single placental gene repeat (PGR) unit encompassing hCS-A and its adjacent 3′-enhancer and 5′ P-element.