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Integration of PHB resulted in lower DNA methylation of the flanking HPRT promoter in females, suggesting the action of a dominant cis-acting escape element.
Given that we observed only one such escape element in 47 genes and 1.5 Mb of DNA, our data support the existence of relatively rare dominant escape elements.
We propose that there is a dominant cis-acting escape element near the PHB gene that allows it to escape from XCI in an otherwise inactivated region on the X chromosome.
The observed reduction of HPRT DNA methylation to 41% on the Xi when adjacent to PHB, from an average of 70% DNA methylation on the Xi for other MaxiP constructs, suggests that the dominant escape element also influenced the HPRT locus.
We determined that the PHB gene escapes from XCI, and as waystations are reduced in abundance on autosomes and NGFR is subject to XCI while being farther from the mouse X-linked DNA than PHB (see Table S2), we conclude that the PHB region likely carries an escape element to escape from XCI.
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Therefore, we hypothesize that there are both waystations and escape elements regulating the spread of XCI.
Escape elements are presumably outside the promoter as none of the 46 MiniPs examined in this study nor the majority of previously examined transgenes escape from XCI.
Models have theorised that there would be waystations, boundary and escape elements involved in XCI (Fig. 2C; reviewed in [ 94]), and to date, multiple elements have been correlated with either genes that escape from or are subject to XCI.
It has been hypothesized that there are domains on the Xi with coordinately regulated XCI caused by nearby XCI way stations spreading XCI or escape elements promoting euchromatin with boundaries separating the two [ 46– 46].
The escaped element that would replicate in the cytoplasm using the ancestral Polinton pPolB spawned two groups of mobile elements (Fig. 2), namely cytoplasmic plasmids (so far found only in fungi) and the " Megavirales," which share the unique trifunctional capping enzyme, RNAP, and D11-like helicase (Fig. 1).
As escape from XCI is frequent in X;autosome translocations [∼30% (reviewed in Yang et al. 2011)], it is likely that, as previously proposed, such escape more generally reflects a lack of waystations rather than the presence of escape elements.
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