What makes the maternal X chromosome

resistant to undergoing imprinted X

rstb.royalsocietypublishing.org inactivation?
Takashi Sado
Department of Bioscience, Graduate School of Agriculture, Kindai University, 3327-204, Nakamachi,
Review Nara 631-8505, Japan

Cite this article: Sado T. 2017 What makes TS, 0000-0002-1232-0250

the maternal X chromosome resistant to
In the mouse, while either X chromosome is chosen for inactivation in
undergoing imprinted X inactivation?
a random fashion in the embryonic tissue, the paternally derived X chromo-
Phil. Trans. R. Soc. B 372: 20160365. some is preferentially inactivated in the extraembryonic tissues. It has been
Downloaded from https://royalsocietypublishing.org/ on 21 November 2022

http://dx.doi.org/10.1098/rstb.2016.0365 shown that the maternal X chromosome is imprinted so as not to undergo inac-
tivation in the extraembryonic tissues. X-linked noncoding Xist RNA becomes
Accepted: 24 April 2017 upregulated on the X chromosome that is to be inactivated. An antisense non-
coding RNA, Tsix, which occurs at the Xist locus and has been shown to
negatively regulate Xist expression in cis, is imprinted to be expressed from
One contribution of 12 to a discussion meeting the maternal X in the extraembryonic tissues. Although Tsix appears to be
issue ‘X-chromosome inactivation: a tribute to responsible for the imprint laid on the maternal X, those who disagree with
Mary Lyon’. this idea would point out the fact that Tsix has not yet been expressed from
the maternal X when Xist becomes upregulated on the paternal but not the
maternal X at the onset of imprinted X-inactivation in preimplantation
Subject Areas: embryos. Recent studies have demonstrated, however, that there is a promi-
genetics nent difference in the chromatin structure at the Xist locus depending on the
parental origin, which I suggest might account for the repression of maternal
Keywords: Xist in the absence of maternal Tsix at the preimplantation stages.
This article is part of the themed issue ‘X-chromosome inactivation:
X chromosome inactivation, imprint,
a tribute to Mary Lyon’.
chromatin structure

Author for correspondence:

Takashi Sado 1. Imprinted and random X inactivation in the mouse
e-mail: tsado@nara.kindai.ac.jp X inactivation is one of the most important epigenetic gene regulations that
operate in early embryogenesis in female mammals [1] and its defect results
in embryonic lethality at the periimplantation stages [2]. It has been assumed
that X inactivation and cellular differentiation are closely linked and mutually
affect their respective processes [3]. There are two waves of X inactivation in the
mouse. In the extraembryonic lineages giving rise to the future placenta and
some extraembryonic membranes, the paternally derived X chromosome is pre-
ferentially inactivated [4], whereas in the embryonic lineage, which will
produce all tissues of the fetus, either of the two X chromosomes is inactivated
in a random fashion regardless of the parental origin. X inactivation first takes
place around the 4–8 cell stages in all the blastomeres in favour of the paternal
X [5,6]. By the time the embryos have developed to the blastocyst stage, the tro-
phectoderm has differentiated to form the outer layer of a blastocyst with the
undifferentiated inner cell mass (ICM) cells inside, a part of which differentiate
into the primitive endoderm 1 day later. While imprinted paternal X inactivation
is maintained in the trophectoderm and primitive endoderm, the inactive
paternal X tentatively restores its transcriptional activity in the remaining ICM
cells that form the epiblast [5,7]. Subsequently, random X inactivation occurs as
the epiblast cells differentiate. Several lines of evidence suggest that the paternal
X is preferentially inactivated due to the presence of an imprint laid on the
maternal X [8–13], which makes it extremely resistant against chromosome silen-
cing, although the possible presence of an imprint laid on the paternal X has not
been completely excluded. An X-linked long noncoding Xist (X inactive-specific

& 2017 The Author(s) Published by the Royal Society. All rights reserved.
 50 kb
Linx Xist Ftx
cen tel
Xite Tsix Jpx/Enox
Xist-AR
Figure 1. Positive regulation of Xist and Tsix by noncoding RNAs mapped in
the X inactivation centre region. Noncoding RNAs transcribed in the same
direction as Xist are delineated on the top and those transcribed in the
same direction as Tsix are at the bottom of the line representing the chro-
mosomal DNA. Centromere (cen) on the left and telomere (tel) on the right.
(Online version in colour.)
transcript) RNA plays a pivotal role in both imprinted and
random X inactivation. Xist RNA is about 18 kb in length,
and becomes upregulated on the future inactive X chromosome
Downloaded from https://royalsocietypublishing.org/ on 21 November 2022
and coats it in cis, probably to form a platform for proteins
involved in heterochromatinization and gene silencing.
Targeted disruption of Xist clearly demonstrated that Xist
is essential for X inactivation to occur in cis, and therefore
an X chromosome deficient for Xist does not undergo X
inactivation [2].
2. Noncoding RNAs in the X inactivation centre
The Xist gene is mapped on the X chromosome in the X inac-
tivation centre (Xic/XIC), which was identified as a master
controlling region required for the X chromosome to be inac-
tivated by classic cytogenetic studies using translocations
between an X chromosome and an autosome [14,15]. There
are several noncoding RNAs transcribed in Xic besides Xist,
which include Jpx/Enox, Ftx, and Xist-AR (figure 1 and
table 1). Jpx transcribed 10 kb upstream of Xist in an opposite
direction relative to Xist has been reported to interact with
and evict CTCF bound at the promoter region of Xist and
consequently to activate transcription of Xist [16]. Ftx was
also suggested to positively regulate Xist via a DNA methyl-
ation-mediated mechanism at the Xist promoter [17] although
it was shown afterward that Ftx is dispensable for imprinted
X inactivation [18]. Xist-AR was recently identified as an anti-
sense RNA occurring at the 50 region of Xist exon1, and was
suggested to upregulate Xist transcription [19]. In addition to
these noncoding RNAs involved in Xist activation, Tsix has
been identified as an RNA that is antisense to Xist and that
covers the entire transcription unit of Xist and negatively
regulates Xist in cis [22–24]. The transcription of Tsix is regu-
lated by the Xite locus producing an enhancer RNA encoded
upstream of the major promoter of Tsix [20] (figure 1).
Another recently identified noncoding RNA, Linx, located
100 kb upstream of Xite, has been suggested to play a role as
a long-range transcriptional regulator for Tsix [21] (figure 1).
Tsix appears to exert its negative effect on Xist transcription at
the onset of X inactivation when the decision of whether or
not Xist is monoallelically upregulated is made. Xist and Tsix
are reciprocally imprinted in the extraembryonic lineages to
be expressed from the paternal and maternal allele, respectively.
Targeted disruption of Tsix, when maternally inherited, causes
ectopic expression of Xist on the maternal X after implantation
and its subsequent inactivation in the extraembryonic tissues of
both female and male embryos, which never occurs in wild-
type, suggesting that Tsix is required for Xist to stay repressed
on the X that remains active [22,24]. Although it is still unknown
Table 1. List of noncoding RNAs that regulate each of Xist and Tsix at Xic. 2
rstb.royalsocietypublishing.org
positive regulator for Xist
Jpx/Enox Sun et al. [16]
Ftx Chureau et al. [17]; Soma et al. [18]
Xist-AR Sarkar et al. [19]
positive regulator for Tsix
Xite Ogawa et al. [20]
Linx Nora et al. [21]
Phil. Trans. R. Soc. B 372: 20160365
which of Tsix RNA or the action of antisense transcription is
important for Tsix’s function, it has been shown that transcrip-
tion of Tsix has to cover the promoter region of Xist to exert
Tsix’s negative effect on Xist [25].
3. An imprint laid on the maternal X to resist
inactivation
The presence of an imprint that renders the maternal X resist-
ant to undergoing inactivation in the extraembryonic tissues
was originally pointed out by Lyon & Rastan [26] and a
series of studies by Takagi and colleagues experimentally
revealed using embryos with a supernumerary maternal X
chromosome that the X chromosomes derived from the
mother indeed bear such an imprint effective only in the
extraembryonic tissues, while this imprint is apparently
erased or ineffective in the embryonic tissue [8,10 –13]. The
presence of two active X chromosomes results in substantial
loss of the polar trophoblast lineages such as the ectoplacen-
tal cone and extraembryonic ectoderm, suggesting that these
tissues are probably most susceptible to the presence of two
active X chromosomes. In contrast, the visceral endoderm, a
derivative of the primitive endoderm, appears to be less
affected by the presence of two active X chromosomes.
It should be pointed out, however, that a small fraction of
cells in the extraembryonic tissues of parthenogenetic embryos
somehow manage to inactivate one of the two maternal X
chromosomes. This seems to be achieved by ectopic upregula-
tion of Xist on either of the maternally derived X chromosomes
at around the morula/blastocyst stages [27,28]. Although the
timing of Xist upregulation in parthenogenetic embryos is rela-
tively late as compared to the upregulation of the paternal Xist
in normal female embryos, these observations suggest that the
imprint of the maternal X can be overridden in parthenogenetic
embryos. In fertilized embryos, however, this imprint
appeared to be so rigid that the maternal X chromosome
does not undergo inactivation in the extraembryonic lineages
even under circumstances in which the paternal X never
becomes inactivated due to the lack of Xist [2]. Interestingly,
embryonic lethality caused by paternal deficiency of Xist is
sometimes fully rescued if combined simultaneously with
maternal deficiency of Tsix. This results in an inverse situation
of imprinted X inactivation in the extraembryonic tissues,
where the maternal X is inactivated and the paternal X remains
active [24]. This demonstrates that it does not necessarily have
to be the paternal X that undergoes inactivation in the extraem-
bryonic tissues as long as dosage compensation of the X-linked
genes is achieved. It has been shown that the maternal imprint
of the X chromosome is established during the period when the
 primary oocyte grows and increases in volume during prophase
of meiosis I [9]. Although neither X chromosome undergoes
inactivation in the majority of cells in the extraembryonic tissues
of parthenogenetic embryos [10], which contain two sets of the
genome derived from the fully grown oocyte, if they are recon-
stituted using the genome of the nongrowing and fully grown
oocyte, the X chromosome derived from the nongrowing
oocyte undergoes inactivation in the extraembryonic tissues.
This clearly demonstrates that the imprint laid on the maternal
X is acquired during oocyte growth.
It would be reasonable to assume that this imprint prevents
upregulation of maternal Xist in the extraembryonic tissues.
Given the fact that the strict repression of maternal Xist is abol-
ished if Tsix is disrupted on the maternal X [22,24], it seems
likely that the maternal expression of Tsix is responsible for pre-
venting upregulation of Xist on the maternal X, and thereby
mediates the maternal imprint against X inactivation. Those
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who disagree this idea would point out the fact that Tsix
expression has not yet been initiated when only the paternal
Xist becomes preferentially upregulated at the 4- to 8-cell
stages [24], and would therefore claim that Tsix would have
nothing to do with the imprint preventing Xist upregulation
on the maternal X at these stages. A possible difference in the
chromatin structure between the maternal and paternal Xist
locus may, however, account for the monoallelic upregulation
of Xist only on the paternal X at these stages. Since the sperm
genome undergoes global exchange from protamine to histone
and becomes decondensed in the zygote, there would be some
differences in the chromatin structure, and therefore in the
accessibility of transcription machinery, between the maternal
and paternal genome. It is possible that these differences cause
a bias in the timing when a subset of genes starts to be
expressed in the early preimplantation embryos in favour of
the paternal alleles. Although a recent study demonstrated
that such parental biases in the timing of gene activation
were not evident at the early preimplantation stages [29], this
does not necessarily rule out the existence of rare loci that
behave in that way. Since Tsix expression initiates on the
maternal X at around the morula stages, if the influence of
differences in the chromatin structure between the two parental
Xist alleles lasts for the first three cell divisions or so, maternal
Xist would stay repressed without Tsix during the early preim-
plantation stages. Tsix upregulated on the maternal X by the
time when the chromatin structure at the Xist locus becomes
equivalent between the paternal and maternal X via multiple
rounds of DNA replication would, in turn, take over the role
in preventing the upregulation of Xist on the maternal X in
the cells that contribute to the extraembryonic lineages.
4. Maternal Xist may simply not be ready for
transcription in the early preimplantation
embryo
A recent study by Fukuda et al. showed that overexpression of
Kdm4b histone H3K9me3 demethylase [30] by introduction of
its mRNA in the oocyte prior to parthenogenetic activation
results in upregulation of Xist at the 4-cell cell stage, which is
rather early for parthenogenetic embryos [31]. This experiment
suggests that decondensation of the maternal genome by
demethylation of H3K9me3 facilitates the expression of Xist
on the maternally derived X chromosomes. They further
showed by DNA-FISH that the chromosome region spanning 3
the Xist locus becomes condensed during the growth of oocytes
rstb.royalsocietypublishing.org
at the prophase of meiosis I, when the maternal imprint against
X inactivation is established [32,33]. Although the condensed
maternal Xist locus relaxes during early preimplantation devel-
opment, the extent of the decondensation across the Xist loci
derived from normally developed oocytes is significantly
smaller than the extents across the Xist loci derived from non-
growing oocytes as well as those on the paternal X in
androgenetic embryos. These studies support the idea that
the Xist locus on the maternal X is more condensed than that
on the paternal X in normal embryos and may need a few
Phil. Trans. R. Soc. B 372: 20160365
rounds of DNA replication to become permissive to transcrip-
tion. Recent ultra-sensitive ChIP-seq analysis revealed
widespread and relatively long-lasting parent-of-origin
asymmetries in preimplantation mouse embryos [34,35].
This may account for our recent finding that the XistCAG
allele, which is generated by replacing the endogenous Xist
promoter with a heterologous CAG promoter to facilitate its
selective monoallelic expression in many types of cells, exhibits
a parent-of-origin-specific difference in the timing of its upre-
gulation in the preimplantation embryos [36]. XistCAG starts
to be expressed from the 4- to 8-cell stages upon paternal trans-
mission, whereas it does not become upregulated until the
blastocyst stage upon maternal transmission. The differential
behaviour of the CAG promoter introduced at the Xist locus
could possibly be due to the difference in the chromatin struc-
ture depending on the parental origin. The CAG promoter in
this context appears to be controlled in the same way as the
endogenous Xist promoter in the early preimplantation
stages. It is tempting to speculate that the difference in the
chromatin structure at the Xist promoter between the paternal
and maternal X underlies the differential behaviour of
the Xist allele according to the parental origin during the
preimplantation development. Given the finding that the
endogenous Xist promoter is enriched in H3K9me3 when
transmitted to the zygotes [32], it is likely that the CAG promo-
ter is also modified in the same manner as the endogenous one
during oogenesis. If this is the case, it raises the question of how
the promoter region of Xist becomes targeted by the machinery
for the repressive modifications such as H3K9me3 in a
sequence-independent manner.
5. Concluding remarks
The presence of an imprint laid on the maternal X chromosome
was predicted based on the cytogenetic observation by Takagi
and colleagues that when multiple maternal X chromosomes
are present, they do not undergo inactivation in the extraem-
bryonic tissues of the early postimplantation embryo. This
imprint is established in the primary oocytes while they are
growing in prophase of meiosis I. Since functional loss of
Tsix results in ectopic inactivation of the maternal X, it is
likely that the imprinted maternal expression of Tsix underlies
the molecular basis for the imprint, which makes the maternal
X resistant against inactivation in the extraembryonic lineages.
I propose that the mechanisms underlying the resistance of the
maternal X against inactivation in the extraembryonic tissues of
the postimplantation embryo pointed out by Takagi and col-
leagues should be distinguished from those that operate at
the preimplantation stages, when only the paternal but not
maternal Xist is expressed. Available evidence suggests that
 the chromatin structure at the Xist locus is different between of imprinted expression of Xist in the preimplantation embryos 4
the paternal and maternal X chromosomes during very early in non-rodents [37] may be related to the fact that they undergo

stages of preimplantation development. I envision that such
a difference in the chromatin structure allows the imprinted
paternal expression of Xist in the preimplantation embryos
while Tsix has not yet been expressed. Although the chromatin
structure of the Xist locus would become equivalent on the
maternal X and paternal X through multiple rounds of DNA
replication, the imprinted expression of Tsix occurring on the
maternal X by then should take over the role of keeping Xist
repressed afterward. It is tempting to speculate that the lack
rstb.royalsocietypublishing.org
more cell division or DNA replication by the time XIST starts
to be expressed [38].
Data accessibility. This article has no additional data.
Competing interests. I declare I have no competing interests.
Funding. T.S. is funded by a Grant-in-Aid for Scientific Research on
Innovative Areas from the Ministry of Education, Science, Sports,
and Culture of Japan (16H01320, 17H05606).

Phil. Trans. R. Soc. B 372: 20160365

References
1. Lyon MF. 1961 Gene action in the X-chromosome 12. Takagi N, Abe K. 1990 Detrimental effects of two 25. Ohhata T, Hoki Y, Sasaki H, Sado T. 2008 Crucial role
Downloaded from https://royalsocietypublishing.org/ on 21 November 2022

of the mouse (Mus musculus L.). Nature 190,
372–373. (doi:10.1038/190372a0)
2. Marahrens Y, Panning B, Dausman J, Strauss W,
Jaenisch R. 1997 Xist-deficient mice are defective in
dosage compensation but not spermatogenesis.
Genes Dev. 11, 156– 166. (doi:10.1101/gad.
11.2.156)
3. Monk M, Harper MI. 1979 Sequential X
chromosome inactivation coupled with cellular
differentiation in early mouse embryos. Nature 281,
311–313. (doi:10.1038/281311a0)
4. Takagi N, Sasaki M. 1975 Preferential inactivation of
the paternally derived X chromosome in the
extraembryonic membranes of the mouse. Nature
256, 640–642. (doi:10.1038/256640a0)
5. Okamoto I, Otte AP, Allis DC, Reinberg D, Heard E.
2004 Epigenetic dynamics of imprinted X
inactivation during early mouse development.
Science 303, 644–649. (doi:10.1126/science.
1092727)
6. Huynh KD, Lee JT. 2003 Inheritance of a pre-
inactivated paternal X chromosome in early mouse
embryos. Nature 426, 857–862. (doi:10.1038/
nature02222)
7. Mak W, Nesterova TB, de Napoles M, Appanah R,
Yamanaka S, Otte AP, Brockdorff N. 2004
Reactivation of the paternal X chromosome in early
mouse embryos. Science 303, 666–669. (doi:10.
1126/science.1092674)
8. Goto Y, Takagi N. 1998 Tetraploid embryos rescue
embryonic lethality caused by an additional
maternally inherited X chromosome in the mouse.
Development 125, 3353 –3363.
9. Tada T, Obata Y, Tada M, Goto Y, Nakatsuji N, Tan S,
Kono T, Takagi N. 2000 Imprint switching for non-
random X-chromosome inactivation during mouse
oocyte growth. Development 127, 3101 –3105.
10. Tada T, Takagi N. 1992 Early development and X-
chromosome inactivation in mouse parthenogenetic
embryos. Mol. Reprod. Dev. 31, 20– 27. (doi:10.
1002/mrd.1080310105)
11. Tada T, Takagi N, Adler ID. 1993 Parental imprinting
on the mouse X chromosome: effects on the early
development of X0, XXY and XXX embryos. Genet.
Res. 62, 139 –148. (doi:10.1017/S001667230
0031736)
active X chromosomes on early mouse
development. Development 109, 189 –201.
13. Shao C, Takagi N. 1990 An extra maternally derived
X chromosome is deleterious to early mouse
development. Development 110, 969 –975.
14. Rastan S, Robertson EJ. 1985 X-chromosome
deletions in embryo-derived (EK) cell lines
associated with lack of X-chromosome inactivation.
J. Embryol. Exp. Morphol. 90, 379– 388.
15. Rastan S. 1983 Non-random X-chromosome
inactivation in mouse X-autosome translocation
embryos—location of the inactivation centre.
J. Embryol. Exp. Morphol. 78, 1 –22.
16. Sun S, Del Rosario BC, Szanto A, Ogawa Y, Jeon Y,
Lee JT. 2013 Jpx RNA activates Xist by evicting
CTCF. Cell 153, 1537–1551. (doi:10.1016/j.cell.
2013.05.028)
17. Chureau C, Chantalat S, Romito A, Galvani A, Duret
L, Avner P, Rougeulle C. 2011 Ftx is a non-coding
RNA which affects Xist expression and chromatin
structure within the X-inactivation center region.
Hum. Mol. Genet. 20, 705–718. (doi:10.1093/hmg/
ddq516)
18. Soma M, Fujihara Y, Okabe M, Ishino F, Kobayashi S.
2014 Ftx is dispensable for imprinted X-
chromosome inactivation in preimplantation mouse
embryos. Sci. Rep. 4, 5181. (doi:10.1038/srep05181)
19. Sarkar MK et al. 2015 An Xist-activating antisense
RNA required for X-chromosome inactivation. Nat.
Commun. 6, 8564. (doi:10.1038/ncomms9564)
20. Ogawa Y, Lee JT. 2003 Xite, X-inactivation intergenic
transcription elements that regulate the probability
of choice. Mol. Cell 11, 731–743. (doi:10.1016/
S1097-2765(03)00063-7)
21. Nora EP et al. 2012 Spatial partitioning of the
regulatory landscape of the X-inactivation centre.
Nature 485, 381–385. (doi:10.1038/nature11049)
22. Lee JT. 2000 Disruption of imprinted X inactivation
by parent-of-origin effects at Tsix. Cell 103, 17 –27.
(doi:10.1016/S0092-8674(00)00101-X)
23. Lee JT, Davidow LS, Warshawsky D. 1999 Tsix, a
gene antisense to Xist at the X-inactivation centre.
Nat. Genet. 21, 400 –404. (doi:10.1038/7734)
24. Sado T, Wang Z, Sasaki H, Li E. 2001 Regulation of
imprinted X-chromosome inactivation in mice
by Tsix. Development 128, 1275–1286.
of antisense transcription across the Xist promoter in
Tsix-mediated Xist chromatin modification.
Development 135, 227 –235. (doi:10.1242/
dev.008490)
26. Lyon MF, Rastan S. 1984 Parental source of
chromosome imprinting and its relevance for
X chromosome inactivation. Differentiation
26, 63 – 67. (doi:10.1111/j.1432-0436.1984.
tb01375.x)
27. Nesterova TB, Barton SC, Surani MA, Brockdorff N.
2001 Loss of Xist imprinting in diploid
parthenogenetic preimplantation embryos. Dev.
Biol. 235, 343–350 (doi:10.1006/dbio.2001.0295)
28. Matsui J, Goto Y, Takagi N. 2001 Control of Xist
expression for imprinted and random X
chromosome inactivation in mice. Hum. Mol. Genet.
10, 1393–1401. (doi:10.1093/hmg/10.13.1393)
29. Deng Q, Ramsköld D, Reinius B, Sandberg R. 2014
Single-cell RNA-seq reveals dynamic, random
monoallelic gene expression in mammalian cells.
Science 343, 193–196. (doi:10.1126/science.
1245316)
30. Fodor BD et al. 2006 Jmjd2b antagonizes H3K9
trimethylation at pericentric heterochromatin in
mammalian cells. Genes Dev. 20, 1557–1562.
(doi:10.1101/gad.388206)
31. Fukuda A, Tomikawa J, Miura T, Hata K,
Nakabayashi K, Eggan K, Akutsu H, Umezawa A.
2014 The role of maternal-specific H3K9me3
modification in establishing imprinted X-
chromosome inactivation and embryogenesis in
mice. Nat. Commun. 5, 5464. (doi:10.1038/
ncomms6464)
32. Fukuda A, Mitani A, Miyashita T, Umezawa A,
Akutsu H. 2015 Chromatin condensation of Xist
genomic loci during oogenesis in mice. Development
142, 4049– 4055. (doi:10.1242/dev.127308)
33. Fukuda A, Mitani A, Miyashita T, Sado T, Umezawa
A, Akutsu H. 2016 Maintenance of Xist imprinting
depends on chromatin condensation state and
Rnf12 dosage in mice. PLoS Genet. 12, e1006375.
(doi:10.1371/journal.pgen.1006375)
34. Zhang B et al. 2016 Allelic reprogramming of the
histone modification H3K4me3 in early mammalian
development. Nature 537, 553– 557. (doi:10.1038/
nature19361)
 35. Zheng H, Huang B, Zhang B, Xiang Y, Du Z, Xu Q,
Li Y. 2016 Resetting epigenetic memory by
reprogramming of histone modifications in
mammals. Mol. Cell 63, 1066–1079. (doi:10.1016/j.
molcel.2016.08.032)
36. Amakawa Y, Sakata Y, Hoki Y, Arata S, Shioda S,
Fukagawa T, Sasaki H, Sado T. 2015 A new Xist
Downloaded from https://royalsocietypublishing.org/ on 21 November 2022
allele driven by a constitutively active promoter
is dominated by Xist locus environment and
exhibits the parent-of-origin effects.
Development 142, 4299–4308. (doi:10.1242/
dev.128819)
37. Okamoto I et al. 2011 Eutherian mammals
use diverse strategies to initiate X-chromosome
inactivation during development. 5
Nature 472, 370– 374. (doi:10.1038/
rstb.royalsocietypublishing.org
nature09872)
38. Sado T, Sakaguchi T. 2013 Species-specific
differences in X chromosome inactivation in
mammals. Reproduction 146, 9. (doi:10.1530/
REP-13-0173)
Phil. Trans. R. Soc. B 372: 20160365