Screen for reactivation of MeCP2 on the inactive X chromosome identifies the BMP/TGF-β superfamily as a regulator of XIST expression

Sripathy, Smitha; Leko, Vid; Adrianse, Robin L.; Loe, Taylor K.; Foss, Eric J.; Dalrymple, Emily; Lao, Uyen; Gatbonton-Schwager, Tonibelle; Carter, Kelly; Payer, Bernhard; Paddison, Patrick J.; Grady, William M.; Lee, Jeannie T.; Bartolomei, Marisa S.; Bedalov, Antonio

doi:10.1073/pnas.1621356114

Cited by 59 publications

(77 citation statements)

References 52 publications

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“…Although no gain-offunction screen has been done to investigate XCI, recently several loss-of-function screens have dissected the mechanism of X chromosome silencing [ Figure 2]. Many of these large-scale screens used a mouse fibroblast cell lines carrying transgenic reporter on Xi (GFP [32,33,40] and luciferase [34] ), to determine the efficiency of Xi reactivation. Quite a few screens identified XCIFs that overlap between these screens, thereby validating the findings and increasing confidence in the RNAi-based screening tools [32][33][34] .…”

Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

“…Many of these large-scale screens used a mouse fibroblast cell lines carrying transgenic reporter on Xi (GFP [32,33,40] and luciferase [34] ), to determine the efficiency of Xi reactivation. Quite a few screens identified XCIFs that overlap between these screens, thereby validating the findings and increasing confidence in the RNAi-based screening tools [32][33][34] . The success of these initial [32][33][34][40][41][42][43] and three proteomics screens [47][48][49] identified more than 500 XCIFs.…”

Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

“…Validation methods qRT-PCR for X-linked genes [40,49] ; RNA-FISH for X-linked genes [32,42,47] ; Xi-Luciferase [34,41] ; Xi-Gfp [33,48] , Immunostaining for H4Kac [43] ; Immunostaining for H3K27me3 [47,48] Biological systems Xi-Gfp [32,33,40] Xi-Luciferase [41] Ch17-TetO-Xist-Mylc2b-Gfp [42] Xi-Mecp2-Hygromycin [34] Ch6-TetO-Xist survival [43] Number of screens performed: screens also encouraged the use of more focused libraries, such as the chromatin modifiers [41] and RNAbinding proteins [42] . While shRNA-based libraries have immensely accelerated the field, use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based libraries is expected to fast-track our understanding of the mechanisms of XCI.…”

Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

“…Previously, XCI has been considered unidirectional during later stages of development, but more recently, the idea of reactivating Xi has emerged. Evidence suggests that the epigenetic state of Xi can be reversed by genetic or pharmacological manipulations [32][33][34] . Although these results were paradigm shifting, they were not unexpected due to the inherent ability of Xi to shuttle between the active and inactive states during embryogenesis.…”

Section: Introductionmentioning

confidence: 99%

“…The reversal of XCI could compensate for the gene deficiencies by expressing the endogenous wild-type copy from Xi. Several studies have demonstrated that upon the interference with the function of regulatory factors of XCI, the Xi can be reactivated in mouse embryonic fibroblasts, fibroblast cell lines and cortical neurons [32][33][34] . However, before embarking on a translational approach based on the idea of X chromosome reactivation (XCR), a detailed understanding of the XCR mechanism and regulatory factors is warranted.…”

Section: Introductionmentioning

confidence: 99%

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Novel molecular players of X chromosome inactivation: new technologies and new insights

Przanowski¹,

Wasko²,

Bhatnagar³

2018

JTGG

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The dosage compensation in placental mammals is achieved by silencing of one copy of the X chromosomes in a female cell by a process called X chromosome inactivation (XCI). XCI ensures equal gene dosage for X-linked genes between the two genders. Although the choice of X chromosome to be silenced is random, once the silencing of the X chromosome has been established, this process is highly regulated and maintained throughout subsequent cell divisions. A long non-coding RNA, Xist , and its interacting proteins execute this multistep process, but several of these regulatory proteins remain unidentified. Recent technological advances based on the genetic and proteomics screening have identified several new regulatory factors as well as dissected the molecular details of XCI regulation. Moreover, identification of regulators of XCI offers an opportunity to explore reactivation of the inactive X chromosome (Xi ) as a potential therapeutic strategy to treat X-linked diseases, like Rett syndrome. Here, we summarize recent reports that identified new regulatory proteins and RNA species playing a crucial role in Xist localization and spreading, recruitment of silencing machinery to the Xi , Xist interaction with chromatin, and structural organization of the Xi in the nuclei.

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Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

Section: Loss-of-function Genetic Screensmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Novel molecular players of X chromosome inactivation: new technologies and new insights

Przanowski¹,

Wasko²,

Bhatnagar³

2018

JTGG

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Sex and the basal mRNA synthesis machinery

et al. 2022

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Evolution and change generated an incredible diversity of organisms on this earth. Yet, some processes are so central to life that change is strongly selected against. Synthesis of the eukaryotic messenger RNA is one example. The assemblies that carry out transcription and processing (capping, polyadenylation, and splicing) are so conserved that most genes have recognizable orthologs in yeast and humans. Naturally, most would conclude transcription and processing are identical in both sexes. However, this is an assumption.Men and women vastly differ in their physiologies. The incidence of pathologies, symptom presentation, disease outcome, and therapeutic response in each sex vary enormously. Despite the harm ignorance causes women, biological research has been historically carried out without regard to sex. The male mouse was the default mammal. A cultured cell's sex was considered irrelevant. Attempts to fill this knowledge gap have revealed molecular dissimilarities. For example, the earliest embryonic male and female transcriptomes differ long before fetal sex hormones appear. We used public data to challenge the assumption of sameness by reviewing reports of sex-biased gene expression and gene targeting. We focused on 120 genes encoding nonregulatory proteins involved in mRNA synthesis. Remarkably, genes with recognizable orthologs in yeast and thus LEAST likely to differ, did differ between the sexes. The rapidly growing public databases can be used to compare the expression of any gene in male and female tissues. Appreciating the principles that drive sex differences will enrich our understanding of RNA biology in all humans-men and women.

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Treating Rett syndrome: from mouse models to human therapies

Vashi

Justice

2019

Mamm Genome

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Rare diseases are very difficult to study mechanistically and to develop therapies for because of the scarcity of patients. Here, the rare neuro-metabolic disorder Rett syndrome (RTT) is discussed as a prototype for precision medicine, demonstrating how mouse models have led to an understanding of the development of symptoms. RTT is caused by mutations in the X-linked gene methyl-CpG-binding protein 2 ( MECP2 ). Mecp2- mutant mice are being used in preclinical studies that target the MECP2 gene directly, or its downstream pathways. Importantly, this work may improve the health of RTT patients. Clinical presentation may vary widely among individuals based on their mutation, but also because of the degree of X chromosome inactivation and the presence of modifier genes. Because it is a complex disorder involving many organ systems, it is likely that recovery of RTT patients will involve a combination of treatments. Precision medicine is warranted to provide the best efficacy to individually treat RTT patients.

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Screen for reactivation of MeCP2 on the inactive X chromosome identifies the BMP/TGF-β superfamily as a regulator of XIST expression

Cited by 59 publications

References 52 publications

Novel molecular players of X chromosome inactivation: new technologies and new insights

Novel molecular players of X chromosome inactivation: new technologies and new insights

Sex and the basal mRNA synthesis machinery

Treating Rett syndrome: from mouse models to human therapies

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