Generation of Cdx2-mCherry knock-in murine ES cell line to model trophectoderm and intestinal lineage differentiation

Jana, Debabrata; Kale, Hanuman T; Shekar, P Chandra

doi:10.1016/j.scr.2019.101521

Cited by 9 publications

(13 citation statements)

References 4 publications

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“…Among different cell types, enterocytes make up the most abundant cell types followed by goblet and enteroendocrine cells which constitute ∼5% and ∼1% of the total epithelial cells, respectively. Among the transcription factors, Cdx2 plays a central role where it was reported that ectopic expression of Cdx2 gene induces intestinal epithelial cell differentiation [ 30 ]. Though several signaling pathways play important roles in patterning gut development and in regulating epithelial differentiation, defects in any of them are linked to many intestinal disorders including frequent intestinal infection, IBD, and celiac disease.…”

Section: Epithelial Functions Of Jak3mentioning

confidence: 99%

Mucosal Epithelial Jak Kinases in Health and Diseases

Kumar

Kuang

Villa

et al. 2021

Mediators of Inflammation

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Janus kinases (Jaks) are a family of nonreceptor tyrosine kinase that include four different members, viz., Jak1, Jak2, Jak3, and Tyk2. Jaks play critical roles in immune cells functions; however, recent studies suggest they also play essential roles in nonimmune cell physiology. This review highlights the significance of epithelial Jaks in understanding the molecular basis of some of the diseases through regulation of epithelial-mesenchymal transition, cell survival, cell growth, development, and differentiation. Growth factors and cytokines produced by the cells of hematopoietic origin use Jak kinases for signal transduction in both immune and nonimmune cells. Among Jaks, Jak3 is widely expressed in both immune cells and in intestinal epithelial cells (IECs) of both humans and mice. Mutations that abrogate Jak3 functions cause an autosomal severe combined immunodeficiency disease (SCID) while activating Jak3 mutations lead to the development of hematologic and epithelial cancers. A selective Jak3 inhibitor CP-690550 (Xeljanz) approved by the FDA for certain chronic inflammatory conditions demonstrates immunosuppressive activity in rheumatoid arthritis, psoriasis, and organ transplant rejection. Here, we also focus on the consequences of Jak3-directed drugs on adverse effects in light of recent discoveries in mucosal epithelial functions of Jak3 with some information on other Jaks. Lastly, we brief on structural implications of Jak3 domains beyond the immune cells. As information about the roles of Jak3 in gastrointestinal functions and associated diseases are only just emerging, in the review, we summarize its implications in gastrointestinal wound repair, inflammatory bowel disease, obesity-associated metabolic syndrome, and epithelial cancers. Lastly, we shed lights on identifying potential novel targets in developing therapeutic interventions of diseases associated with dysfunctional IEC.

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Section: Epithelial Functions Of Jak3mentioning

confidence: 99%

Mucosal Epithelial Jak Kinases in Health and Diseases

Kumar

Kuang

Villa

et al. 2021

Mediators of Inflammation

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“…In this study, we systematically sought species specific mutations in the DBD of TFs and identified Cdx2 as an apparent candidate for adaptive evolution in the mouse lineage with three specific amino acid changes exclusively present in mCdx2 DBD. Given that Cdx2 plays an important role in embryonic development and stem cell differentiation [31,32,37], we investigated its function in mESC differentiation. In line with previous studies [30], induction of mCdx2 caused dramatic transcriptomic and epigenetic changes, confirming the crucial role of mCdx2 in cell differentiation and lineage specification.…”

Section: Discussionmentioning

confidence: 99%

“…Cdx2 is a homeobox transcription factor essential for the development of the intestinal epithelium and the placenta [32,36]. Serving as the first lineage specification marker [30][31][32]37], previous studies have shown that ectopic overexpression of Cdx2 could efficiently promote the differentiation of ES cells to trophoblast stem cells [30,31,33]. Therefore, to investigate the function of Cdx2 during this process, we applied a doxycycline (DOX) inducible The massive differences in gene expression after mCdx2 induction suggested that dramatic differences in the epigenetic landscapes could be induced after mCdx2 overexpression (mCdx2-OE).…”

Section: Cdx2 Functions As a Pioneer Factor And Induces Trophoblast-like Epigenomic And Transcriptomic Landscapementioning

confidence: 99%

“…The finding was further substantiated by including 56 species as well as 37 mouse strains in the sequence comparison. Given the established function of Cdx2 in lineage specification and trophectoderm differentiation [30][31][32][33], we investigated the potential effect of the three mouse-specific amino acid changes in the mouse embryonic stem cell (mESC) systems. Unexpectedly, we did not observe any significant effects at either DNA binding specificity or target gene expression induced by the three changes.…”

Section: Introductionmentioning

confidence: 99%

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Evolutionary Analysis of Transcriptional Regulation Mediated by Cdx2 in Rodents

Liang

Cui

et al. 2021

Preprint

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Differences in gene expression, which can arise from divergence in cis-regulatory elements or alterations in transcription factors binding specificity, are one of the most important causes of phenotypic diversity during evolution. By protein sequence analysis, we observed high sequence conservation in the DNA binding domain (DBD) of the transcription factor Cdx2 across many vertebrates, whereas three amino acid changes were exclusively found in mouse Cdx2 (mCdx2), suggesting potential positive selection in the mouse lineage. Multi-omics analyses were then carried out to investigate the effects of these changes. Surprisingly, there were no significant functional differences between mCdx2 and its rat homologue (rCdx2), and none of the three amino acid changes had any impact on its function. Finally, we used rat-mouse allodiploid embryonic stem cells (RMES) to study the cis effects of Cdx2-mediated gene regulation between the two rodents. Interestingly, whereas Cdx2 binding is largely divergent between mouse and rat, the transcriptional effect induced by Cdx2 is conserved to a much larger extent.Author summaryOur study 1) represented a first systematic analysis of species-specific adaptation in DNA binding pattern of transcription factor. Although the mouse-specific amino acid changes did not manifest functional impact in our system, several explanations may account for it (See Discussion part for the detail); 2) represented a first study of cis-regulation between two reproductively isolated species by using a novel allodiploid system; 3) demonstrated a higher conservation of transcriptional output than that of DNA binding, suggesting the evolvability/plasticity of the latter; 4) finally provided a rich data resource for Cdx2 mediated regulation, including gene expression, chromatin accessibility and DNA binding etc.

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“…The nding was further substantiated by including 56 species as well as 37 mouse strains in the sequence comparison. Given the established function of Cdx2 in lineage speci cation and trophectoderm differentiation [30][31][32][33], we investigated the potential effect of the three mouse-speci c amino acid changes in the mouse embryonic stem cell (mESC) systems. Unexpectedly, we did not observe any signi cant effects at either DNA binding speci city or target gene expression induced by the three changes.…”

Section: Introductionmentioning

confidence: 99%

Evolutionary Analysis of Transcriptional Regulation Mediated by Cdx2 in Rodents

Liang

Cui

et al. 2021

Preprint

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Background: Differences in gene expression, which arises from divergence in cis-regulatory elements or alterations in transcription factors (TFs) binding specificity, are one of the most important causes of phenotypic diversity during evolution. On one hand, changes in the cis-elements located in the vicinity of target genes affect TF binding and/or local chromatin environment, thereby modulating gene expression in one-to-one manner. On the other hand, alterations in trans-factors influence the expression of their target genes in a more pleiotropic fashion. Although evolution of amino acid sequences is much slower than that of non-coding regulatory elements, particularly for the TF DNA binding domains (DBD)， it is still possible that changes in TF-DBD might have the potential to drive large phenotypic changes if the resulting effects have a net positive effect on the organism’s fitness. If so, species-specific changes in TF-DBD might be positively selected. So far, however, this possibility has been largely unexplored.Results: By protein sequence analysis, we observed high sequence conservation in the DNA binding domain (DBD) of the transcription factor Cdx2 across many vertebrates, whereas three amino acid changes were exclusively found in mouse Cdx2 (mCdx2), suggesting potential positive selection in the mouse lineage. Multi-omics analyses were then carried out to investigate the effects of these changes. Surprisingly, there were no significant functional differences between mCdx2 and its rat homologue (rCdx2), and none of the three amino acid changes had any impact on its function. Finally, we used rat-mouse allodiploid embryonic stem cells (RMES) to study the cis effects of Cdx2-mediated gene regulation between the two rodents. Interestingly, whereas Cdx2 binding is largely divergent between mouse and rat, the transcriptional effect induced by Cdx2 is conserved to a much larger extent.Conclusions: There were no significant functional differences between mCdx2 and its rat homologue (rCdx2), and none of the three amino acid changes had any impact on its function. Moreover, Cdx2 binding is largely divergent between mouse and rat, the transcriptional effect induced by Cdx2 is conserved to a much larger extent.

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Generation of Cdx2-mCherry knock-in murine ES cell line to model trophectoderm and intestinal lineage differentiation

Cited by 9 publications

References 4 publications

Mucosal Epithelial Jak Kinases in Health and Diseases

Mucosal Epithelial Jak Kinases in Health and Diseases

Evolutionary Analysis of Transcriptional Regulation Mediated by Cdx2 in Rodents

Evolutionary Analysis of Transcriptional Regulation Mediated by Cdx2 in Rodents

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