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Dale, R.

Publications and source records attributed to Dale, R..

At least 19 recordsLinked to original sources

WS6 enables scalable ex vivo expansion and gene editing of epithelial basal stem cells

Modeling human epithelial diseases and developing cell-based therapies require robust methods to expand and manipulate epithelial stem and progenitor cells in vitro. Basal stem/progenitor cells from stratified epithelia can be expanded in 3T3-J2 fibroblast feeder cell co-culture systems, and the addition of the ROCK inhibitor Y-27632 enhances proliferation and culture longevity, a phenomenon described as conditional reprogramming. Here, we present a method incorporating the small molecule WS6 to further improve the proliferation and lifespan of cultured epithelial cells from multiple tissues, including airway, skin, and thymus. Cells maintained in this medium ( EpMED; FAD+Y+WS6) retain basal stem/progenitor cell identity and function, including the capacity to differentiate. We demonstrate their capacity to engraft in vivo in a tracheal transplantation model. In a second application, we generate clonal CRISPR-Cas9 genome edited nasal cultures, introducing targeted knockouts of DNAH5 or DNAI2 to create primary ciliary dyskinesia disease models. We anticipate that our method will have broad applications in epithelial cell biology, disease modeling, and regenerative medicine, while reducing reliance on immortalized or cancer cell lines and animal experimentation.

cell biology↗

RNA-RNA Interactome Approaches Provide in vivo Evidence for a Critical Role of the Hfq Rim Face in sRNA-mRNA Pairing

Most bacterial small regulatory RNAs (sRNAs) modulate gene expression by forming complementary base pairs with target mRNAs, dependent upon the RNA chaperone Hfq. Hfq has three RNA-binding faces (proximal, rim, and distal), facilitating simultaneous binding of sRNAs and target mRNAs. Here, we systematically examined the functional impact of point mutations in the RNA binding faces on in vivo pairing, using the RNA Interaction by Ligation and Sequencing (RIL-seq) approach. The distal and proximal Hfq binding face mutants retained substantial numbers of RNA-RNA interactions (significant chimera counts or S-chimeras). However, the rim face mutant R16A showed a near-complete loss of S-chimeras, although Hfq R16A retained partial RNA binding activity as well as partial regulatory activity. Intracellular RIL-seq (iRIL-seq), a method with fewer in vitro processing steps, led to more S-chimeras in R16A but there were still fewer than for wild type Hfq and somewhat different sets of prevalent RNA-RNA pairs. Our analysis provides insights into how the RNA-binding faces of Hfq contribute to pairing in vivo, document the key role for the rim face in stabilizing RNA pairs on Hfq, and highlight intriguing differences captured by different RNA-RNA interactome approaches. GRAPHIC ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/660752v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@99569aorg.highwire.dtl.DTLVardef@104d130org.highwire.dtl.DTLVardef@141b117org.highwire.dtl.DTLVardef@1d19611_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

Making new connections: An fNIRS machine learning classification study of neural synchrony in the default mode network

Successfully making connections with others is crucial to navigating the social world and general well-being, yet little is known about connection formation and its neurocognitive underpinnings. Increasingly, neuroscientists use interpersonal neural synchrony within the default mode network (DMN) to measure when two or more people subjectively experience something in similar ways. DMN synchrony as seeing eye-to-eye is typically observed when multiple people are passively observing the same stimulus. In this study, we tested whether the same DMN synchrony as seeing eye-to-eye pattern holds during social interactions. We conducted a between-subject naturalistic experiment with 70 pairs of strangers engaged in either shallow or deep conversations while brain activity was measured with functional near infrared spectroscopy (fNIRS). Stranger dyads successfully formed connections, as indicated by composite connection scores. Replicating Kardas et al. (2021), those in the deep conversation condition felt more connected than those in the shallow conversation condition. DMN neural synchrony significantly predicted self-reported connection, with synchrony in the DMN subregions of medial prefrontal cortex (mPFC) and right temporoparietal junction (TPJ) each correlating significantly with connection. Using machine learning classification, we distinguished high-versus low-connection dyads based on DMN neural synchrony and the perceived depth of conversation with 64.5% accuracy across 1,000 iterations. This effect was primarily carried by right TPJ, which alone classified connection strength at 62.6% accuracy. We consider implications related to the growing loneliness crisis and the importance of understanding how social connections can be formed and fostered in an era of increased social isolation. Significance StatementThe current loneliness epidemic has serious consequences on health and well-being. Forming interpersonal connections is crucial for alleviating loneliness, yet little is known about its neural basis. Neural synchrony, a potential biological marker of people being on the same page, may be an indicator of social connection. We recorded brain activity as strangers engaged in a get-to-know-you conversation and found that neural synchrony--specifically within the default mode network (DMN) and subregions including medial prefrontal cortex (mPFC), and right temporoparietal junction (TPJ)--predicted self-reported connection. Machine learning accurately classified high- and low-connection dyads based on DMN synchrony and perceived conversation depth. These findings suggest that deeper conversations, though more effortful, may foster stronger social bonds with measurable neural correlates.

neuroscience↗

The Role of SIAMESE in G2 Checkpoint Regulation in Arabidopsis thaliana

In Arabidopsis, SIAMESE (SIM) is a cyclin-dependent kinase inhibitor that restricts progression through mitosis. SIM is well known as a regulator of endoreplication in trichomes and roots. Mathematical modeling of the cell cycle has indicated that SIM may also modulate the length of G2 during mitotic cycles, potentially replacing the WEE1/CDC25 circuit that regulates G2 timing in animals and fungi, which is absent in plants. The predictions of this model were tested in several ways. First, the root meristem is longer in sim mutant roots than in wild-type (WT) roots. Second, two independent methods of measuring cell cycle phases, long-term live-cell imaging using fluorescent protein cell cycle markers and 5-ethynyl-2-deoxyuridine (EdU) pulse-chase imaging, showed that the length of G2 in root meristem cortex cells is shorter in sim mutant plants compared to WT, consistent with the view that these changes in G2 length are due to greater G2 CDK activity in sim mutants. Additionally, the fluorescence of a CYCB:GFP fusion both rose and declined more sharply in the mutant than in wild-type. Because both transcription and degradation of this fusion are directly affected by G2 CDK activity, this result is consistent with sim mutants having greater G2 CDK activity. Taken together, these results suggest that, in addition to its known role in inducing endoreplication, SIM may play a role in regulating the length of G2 during mitotic cycles, at least partially replacing the absence of WEE1/CDC25 regulation of the mitotic cell cycle.

plant biology↗

Reducing methylation of histone 3.3 lysine 4 in the medial ganglionic eminence and hypothalamus recapitulates neurodevelopmental disorder phenotypes

Methylation of lysine 4 on histone H3 (H3K4) is enriched on active promoters and enhancers and correlates with gene activation. Disruption of H3K4 methylation is associated with numerous neurodevelopmental diseases (NDDs) that display intellectual disability and abnormal body growth. Here, we perturb H3K4 methylation in the medial ganglionic eminence (MGE) and the hypothalamus, two brain regions associated with these disease phenotypes. These mutant mice have fewer forebrain interneurons, deficient network rhythmogenesis, and increased spontaneous seizures and seizure susceptibility. Mutant mice are significantly smaller than control littermates, but they eventually became obese due to striking changes in the genetic and cellular hypothalamus environment in these mice. Perturbation of H3K4 methylation in these cells produces deficits in numerous NDD-associated behaviors, with a bias for more severe phenotypes in female mice. Single cell sequencing reveals transcriptional changes in the embryonic and adult brain that underlie many of these phenotypes. In sum, our findings highlight the critical role of H3K4 methylation in regulating survival and cell-specific gene regulatory mechanisms in forebrain GABAergic and hypothalamic cells during neurodevelopment to control network excitability and body size homoeostasis.

neuroscience↗

A novel transgenic reporter to study vertebrate epigenetics

Epigenetic reprogramming contributes to the generation of cellular diversity during vertebrate development but the mechanisms directing this are still not well understood. Large-scale genetic screens have been highly successful in identifying epigenetic regulatory genes in invertebrates such as worms and flies, but similar large-scale genetic screens to identify epigenetic regulators have not been carried out in vertebrates. Here we report a newly generated "EpiTag" zebrafish transgenic reporter line that permits easy cellular-level visualization of epigenetic silencing or activation in living animals during development, gametogenesis, and regeneration. We use the EpiTag reporter to carry out an F3 ENU mutagenesis screen for epigenetic silencing or activating mutants, identifying relevant vertebrate tissue-specific epigenetic regulatory genes including a new epigenetic model for metabolic dysfunction-associated fatty liver disease (MAFLD). The EpiTag reporter line represents a powerful new tool for genetic and experimental analysis of tissue-specific epigenetic gene regulation in vertebrates. One Sentence SummaryEpiTag transgenic zebrafish provide a powerful new tool for visualizing and studying epigenetic regulation in living vertebrate animals.

genomics↗

Early epigenetic priming of regeneration studied with a novel transgenic epigenetic reporter

Tissue regeneration requires previously differentiated cells to regain developmental plasticity. However, the upstream mechanisms initiating this process remain poorly understood. Here, we leverage a novel "EpiTag" transgenic zebrafish reporter line that enables real-time visualization of epigenetic silencing and activation to identify and carry out a comprehensive multi-omics analysis of cells undergoing epigenetic reprogramming during caudal fin regeneration. EpiTag GFP expression is transiently activated in cells contributing to regeneration between 12 to 16 hours post amputation (hpa), preceding the expression of canonical blastema markers. Single-cell RNA-seq reveals that GFP+ cells are restricted to regeneration-competent lineages such as pre-osteoblasts, proliferating cells, and wound epithelium. Integrated bulk RNA-seq, time-course RNA-seq, ATAC-seq, and bisulfite-seq on FACS-isolated GFP+ cells uncovers an early gene expression module enriched for chromatin regulators and a late gene expression module enriched for morphogenesis genes. Chromatin accessibility and DNA methylation changes are strongly associated with these late-expressed genes, suggesting epigenetic priming. We identify a number of epigenetic factors upregulated in the early gene expression module and show that ruvbl1 and ruvbl2, components of ATP-dependent chromatin remodeling complexes, are required for proper regeneration in both adult fins and larval tails. Our results establish EpiTag transgenics as a powerful in vivo tool for studying epigenetic reprogramming and highlight early chromatin remodeling events that enable activation of regenerative gene expression programs.

genomics↗

Young KRAB-zinc finger gene clusters are highly dynamic incubators of ERV-driven genetic heterogeneity in mice

KRAB-zinc finger proteins (KZFPs) comprise the largest family of mammalian transcription factors, rapidly evolving within and between species. Most KZFPs repress endogenous retroviruses (ERVs) and other retrotransposons, with KZFP gene numbers correlating with the ERV load across species, suggesting coevolution. How new KZFPs emerge in response to ERV invasions is currently unknown. Using a combination of long-read sequencing technologies and genome assembly, we present a first detailed comparative analysis of young KZFP gene clusters in the mouse lineage, which has undergone recent KZFP gene expansion and ERV infiltration. Detailed annotation of KZFP genes in a cluster on Mus musculus Chromosome 4 revealed parallel expansion and diversification of this locus in different mouse strains (C57BL/6J, 129S1/SvImJ and CAST/EiJ) and species (Mus spretus and Mus pahari). Our data supports a model by which new ERV integrations within young KZFP gene clusters likely promoted recombination events leading to the emergence of new KZFPs that repress them. At the same time, ERVs also increased their numbers by duplication instead of retrotransposition alone, unraveling a new mechanism for ERV enrichment at these loci.

genetics↗

Multi-scale dynamics influence the division potential of stomatal lineage ground cells in Arabidopsis

During development, many precursor lineages are flexible, producing variable numbers and types of progeny cells. What factors determine whether a precursor cell differentiates or retains the capacity to divide? Here, we leverage the developmental flexibility of the stomatal lineage ground cell (SLGC) in Arabidopsis leaves as a model for how flexible decisions are regulated. Using a quantitative approach that combines long-term live imaging and statistical modeling, we discover that cell size is a strong predictor of SLGC behaviour: larger SLGCs divide less often than smaller cells. We propose that cell size is linked to division behaviour at multiple spatial scales. At the neighbourhood scale, cell size correlates with the strength of cell-cell signaling, which affects the rate at which SPEECHLESS (SPCH), a division-promoting transcription factor, is degraded. At the subcellular scale, cell size correlates with nuclear size, which modulates the concentration of SPCH in the nucleus. Our work shows how initial differences in SPCH levels are canalized by nuclear size and cell-cell signaling to inform the behaviour of a flexible cell type.

developmental biology↗

Mapping the Learning Curves of Deep Learning Networks

There is an important challenge in systematically interpreting the internal representations of deep neural networks. This study introduces a multi-dimensional quantification and visualization approach which can capture two temporal dimensions of a model learning experience: the "information processing trajectory" and the "developmental trajectory." The former represents the influence of incoming signals on an agents decision-making, while the latter conceptualizes the gradual improvement in an agents performance throughout its lifespan. Tracking the learning curves of a DNN enables researchers to explicitly identify the model appropriateness of a given task, examine the properties of the underlying input signals, and assess the models alignment (or lack thereof) with human learning experiences. To illustrate the method, we conducted 750 runs of simulations on two temporal tasks: gesture detection and natural language processing (NLP) classification, showcasing its applicability across a spectrum of deep learning tasks. Based on the quantitative analysis of the learning curves across two distinct datasets, we have identified three insights gained from mapping these curves: nonlinearity, pairwise comparisons, and domain distinctions. We reflect on the theoretical implications of this method for cognitive processing, language models and multimodal representation. Author summaryDeep learning networks, specifically recurrent neural networks (RNNs), are designed for processing incoming signals sequentially, making them intuitive computational systems for studying cognitive processing that involves dynamic contexts. There has been a tradition in the fields of machine learning and neuro-cognitive science to examine how a system (either humans or models) represents information through various computational and statistical techniques. Our study takes this one step further by devising a technique for examining the "learning curves" of deep learning networks utilizing the sequential representations as part of RNNs architectures. Just as humans develop learning curves when solving problems, the introduced method captures both how incoming signals help improve decision-making and how a systems problem-solving abilities enhance when encountering the same situation multiple times throughout its lifespan. Our study selected two distinct tasks: gesture detection and emotion tweet classification, to illustrate the insights researchers can draw from mapping models learning curves. The proposed method hinted that gesture learning experiences are smoother, while language learning relies on sudden knowledge gains during processing, corroborating the findings from previous literature.

scientific communication and education↗

Changes in TP53 gene, telomere length, mitochondrial DNA, and its amount in benign prostatic hyperplasia patients

Benign prostatic hyperplasia (BPH) is a growing issue due to an ageing population; however, there are not enough studies connecting it with ageing hallmarks. Some of the ageing hallmarks are changes in telomere length (TL) and genetic changes like possible mutations in the TP53 gene or mitochondrial genome (mtDNA). Our study investigated these factors to see if they could be linked with BPH. Prostate tissue samples were obtained from 32 patients with BPH (and 30 blood samples). As a healthy control group, age-matching blood DNA samples were used. For mtDNA sequence data comparison, 50 samples of a general Latvian population were used. The full mtDNA genome was sequenced using Next Generation Sequencing (NGS), for TP53 gene - Sanger sequencing, and for mtDNA amount and telomere length - qPCR assay. BPH patients in prostate tissue had much higher TL than in blood cells, and the healthy controls had the shortest telomeres. Also, the mtDNA amount in BPH prostate tissue was the highest compared with blood, and controls had the smallest amount. In the TP53 gene, we did not find any mutations that could be linked to BPH. In the mtDNA genome, we found several unique mutations and heteroplasmic changes, as well as changes that have been linked before with prostate cancer (PC). In conclusion, prolonged telomeres and changes in mtDNA amount might be involved in the development of BPH. Concerning mtDNA genome changes, some of the heteroplasmic or homoplasmic variants might contribute to the development of BPH. More studies should be conducted to prove or disapprove of these connections.

genetics↗

Discovery of active mouse, plant and fungal cytochrome P450s in endogenous proteomes and upon expression in planta.

Eukaryotes produce a large number of cytochrome P450s that mediate the synthesis and degradation of diverse endogenous and exogenous metabolites. Yet, most of these P450s are uncharacterized and global tools to study these challenging, membrane-resident enzymes remain to be exploited. Here, we applied activity profiling of plant, mouse and fungal P450s with chemical probes that become reactive when oxidized by P450 enzymes. Identification by mass spectrometry revealed labeling of a wide range of active P450s, including six plant P450s, 40 mouse P450s and 13 P450s of the fungal wheat pathogen Zymoseptoria tritici. We next used transient expression of GFP-tagged P450s by agroinfiltration to show ER-targeting and NADPH-dependent, activity-based labeling of plant, mouse and fungal P450s. Both global profiling and transient expression can be used to detect a broad range of active P450s to study e.g. their regulation and discover selective inhibitors.

plant biology↗

Competition for resources during development drives allometric patterns in the grass Setaria

Plant growth and resilience is greater than the sum of its component traits, with important traits influencing one another. This interdependency makes it challenging to identify the genetic determinants of key agronomic traits, such as water use efficiency. Measuring traits such as plant height and size is possible via image analysis software, such as PlantCV, but these traits are often highly correlated. Furthermore, plant size is estimated using a 2D projection of a 3D object, which is more difficult for plants with complex body plans. To address these problems, we developed a generalizable, biologically-informed model describing the temporal coordination of semi-sequential phytomeric growth in the model grass Setaria. Our approach integrates time dependence with water usage, the growth of phytomers, and the emergence of side shoots or tillers, improving our ability to estimate both phytomer-level phenotypes and water usage traits in high-throughput. We developed new PlantCV methods to inform the Phytomeric Growth Model, and estimated parameter traits using a high-throughput phenotyping dataset of a recombinant inbred line (RIL) panel in well-watered and drought conditions. Model parameter estimates identified additional quantitative trait loci (QTL) for new traits compared to the directly measured PlantCV-derived estimates, and predicted the relationships underlying QTL control of composite traits through co-localization of model parameters to loci controlling size and water use metrics. The Phytomeric Growth Model estimates improved our ability to estimate the growth of tillers and obtain a model-based estimate of marginal water use efficiency (defined here as the ratio of biomass gained per gram of water transpired), both identified as highly influential on plant drought responses through random forest classification. Our approach demonstrates the value of integrating mechanistic modeling with high-throughput imaging to extract new information at scale.

plant biology↗

Mathematical model of RNA-directed DNA methylation predicts siRNA production in bursts

RNA-directed DNA Methylation (RdDM) is a plant-specific de novo methylation pathway that is responsible for maintenance of asymmetric methylation (CHH, where H=A, T, or G) in euchromatin. Loci with CHH methylation are transcriptionally silent and produce 24-nucleotide (nt) short interfering (si) RNAs. These siRNAs direct additional CHH methylation to the locus, thereby maintaining methylation states through DNA replication. To understand the necessary conditions to produce stable CHH methylation, we developed a stochastic mathematical model of RdDM. The model describes DNA target search of DNA or RNA by siRNAs derived from CHH-methylated loci. When the siRNA (bound by an Argonaute protein) finds the matching locus, the model uses the dwell time of the matched complex to determine the degree of CHH reinforcing methylation. Reinforcing methylation occurs either throughout the cell cycle (steady reinforcement), or immediately following replication (bursty reinforcement). Each simulation occurs over 10 cell cycles, and for 7 simulation replicates. We use nonparametric statistics to compare initial and final CHH methylation distributions to determine whether the simulation conditions produce stable maintenance. We apply this method to the low CHH methylation case, wherein the median is only 8%, and many loci have less than 8% methylation. The resulting model predicts that siRNA production must be linearly proportional to CHH methylation levels at each locus, that bursty reinforcement produces more stable systems, and that slightly higher levels of siRNA production are required for DNA target search, compared to RNA target search. Unlike CG methylation, which typically exhibits bi-modality, with loci having either 100% or 0% methylation, CHH methylation putatively exists at a range of methylation fractions. Our model predicts that careful tuning of the negative feedbacks in the system are required to balance the positive feedback loop of increasing CHH methylation and increasing siRNA production, enabling stable maintenance of a range of CHH methylation across replication events.

genetics↗

A cell size threshold triggers commitment to stomatal fate in Arabidopsis

How flexible developmental programs integrate information from internal and external factors to modulate stem cell behavior is a fundamental question in developmental biology. Cells of the Arabidopsis stomatal lineage modify the balance of stem cell proliferation and differentiation to adjust the size and cell type composition of mature leaves. Here, we report that meristemoids, one type of stomatal lineage stem cell, trigger the transition from asymmetric self-renewing divisions to commitment and terminal differentiation by crossing a critical cell size threshold. Through computational simulation, we demonstrate that this cell size-mediated transition allows robust, yet flexible termination of stem cell proliferation and we observe adjustments in the number of divisions before the differentiation threshold under several genetic manipulations. We experimentally evaluate several mechanisms for cell size sensing, and our data suggest that cell size is sensed via a chromatin ruler acting in the nucleus.

developmental biology↗

Integrating Math Modeling, Coding, and Biology in aCURE Lab

The development of mathematical and quantitative skills is increasingly critical for biology students. Literacy in coding, statistics, and mathematical modeling enables students to engage in systems-thinking and the analysis of large data sets. Here we present a flexible tool illustrating concepts in mathematical modeling, coding, and biology for integration into both traditional cookbook and inquiry-driven labs for freshmen biology students. We developed simple and complex mathematical models of potato catechol oxidase, a popular system to teach enzyme kinetics in undergraduate biology labs. We integrated both models into a freely-available web app for simulation and parameter estimation. The models are usable even if experimental details are unknown or poorly controlled, so that even novice students can work through the problem. We illustrate this by estimating the kinetic parameters of catechol oxidase in the complex model using data obtained from two sections of a course-based undergraduate research experience (CURE) freshman biology lab. Worksheets with questions motivating model building and simulation comprehension are provided. The effect of these exercises on students opinions of math, biology, and coding are evaluated using pre- and post-test surveys and student feedback. Our results show that these tools illustrate enzyme kinetics mathematically, students are not intimidated by the degree of math or coding involved, and in some cases are interested to do more, despite being unaware of the focus of the lab when signing up.

scientific communication and education↗

A dynamic model of the ABA Signaling pathway with its core components: translation rate of PP2C determines the kinetics of ABA-induced gene expression

The abscisic acid (ABA) signaling pathway is the key defense mechanism against drought stress in plants, yet the connectivity of cellular molecules related to gene expression in response to ABA is little understood. A dynamic model of the core components of the ABA signaling pathway was built using ordinary differential equations to understand the connectivity. Parameter values of protein-protein interactions and enzymatic reactions in the model were implemented from the data obtained by previously conducted experiments. On the other hand, parameter values of gene expression and translation were determined by comparing the kinetics of gene expression in the model to those of ABA-induced RD29A (response to desiccation 29A) in actual plants. Based on the analyses of the optimized model, we hypothesized that the translation rate of PP2C (protein phosphatase type 2C) is downregulated by ABA to increase the ABRE (ABA-responsive element) promoter activity. The hypotheses were preliminarily supported by newly conducted experiments using transgenic Arabidopsis plants that carry a luciferase expression cassette driven by the RD29A promoter (RD29A::LUC). The model suggests that identifying a mechanism that alters PP2C translation rate would be one of the next research frontiers in the ABA signaling pathway.

systems biology↗

Rapid redistribution and extensive binding of NANOG and GATA6 at shared regulatory elements underlie specification of divergent cell fates

Establishment of divergent cell types from a common progenitor requires transcription factors (TFs) to promote lineage-restricted transcriptional programs while suppressing alternative fates. In the mouse blastocyst, cells of the inner cell mass (ICM) coexpress NANOG and GATA6, two TFs that drive the bifurcation of these progenitors into either the epiblast (Epi) or the primitive endoderm (PrE), respectively. Here, using in vitro differentiation, we describe the molecular mechanisms of how GATA6 quickly induces the PrE fate while repressing the Epi lineage. GATA6 functions as a pioneer TF by inducing nucleosome repositioning at regulatory elements controlling PrE genes, making them accessible for deposition of active histone marks and leading to rewiring of chromatin interactions and ultimately transcriptional activation. GATA6 also binds most regulatory elements of Epi genes followed by eviction of the Epispecific TFs NANOG and SOX2, loss of active histone marks, and reduction in chromatin accessibility that culminates in transcriptional repression. Unexpectedly, evicted NANOG and SOX2 transiently bind PrE regulatory elements occupied by GATA6. Our study shows that GATA6 binds and modulate the same regulatory elements as Epi TFs, a phenomenon we also validated in blastocysts. We propose that the ability of PrE and Epi-specific TFs to extensively bind and regulate the same gene networks contributes to ICM plasticity and allows rapid cell lineage specification by coordinating both activation

developmental biology↗