Miracle-dz/UniFAR-FTUs huggingface dataset:
Fibroblasts-derived from Pluripotent Cells Harboring a Single Allele Knockout in Two Pluripotency Genes Exhibit DNA Methylation Abnormalities and pluripotency induction Defects

A complete knockout (KO) of a single key pluripotency gene has been shown to drastically affect embryonic stem cell (ESC) function and epigenetic reprogramming. However, knockin (KI)/KO of a reporter gene only in one of two alleles in a single pluripotency gene is considered harmless and is largely used in the stem cell field. Here, we sought to understand the impact of simultaneous elimination of a single allele in two ESC key genes on pluripotency potential and acquisition. We established multiple pluripotency systems harboring KI/KO in a single allele of two different pluripotency genes (i.e. Nanog+/-; Sall4+/-, Nanog+/-; Utf1+/-, Nanog+/-; Esrrb+/- and Sox2+/-; Sall4+/-). Interestingly, although these double heterozygous mutant lines maintain their stemness and contribute to chimeras equally to their parental control cells, fibroblasts derived from these systems show a significant reduction in their capability to induce pluripotency either by Oct4, Sox2, Klf4 and Myc (OSKM) or by nuclear transfer (NT). Tracing the expression of Sall4 and Nanog, as representative key pluripotency targeted genes, at early phases of reprogramming could not explain the seen delay/blockage. Further exploration identifies abnormal methylation landscape around pluripotent and developmental genes in the double heterozygous mutant fibroblasts. Accordingly, treatment with 5-azacytidine two days prior to transgene induction rescues the reprogramming defects. This study emphasizes the importance of maintaining two intact alleles for pluripotency induction and suggests that insufficient levels of key pluripotency genes leads to DNA methylation abnormalities in the derived-somatic cells later on in development.
Root-specific activation of plasma membrane H+-ATPase 1 enhances plant growth and shoot accumulation of nutrient elements under nutrient-poor conditions in Arabidopsis thaliana

Plasma membrane (PM) H+-ATPase contributes to nutrient uptake and stomatal opening by creating proton gradient across the membrane. Previous studies report that a dominant mutation in the OPEN STOMATA2 locus (OST2-2D) constitutively activates Arabidopsis PM H+-ATPase 1 (AHA1), which enlarges proton motive force for root nutrient uptake. However, the stomatal opening is also constitutively enhanced in the ost2-2D, causing the drought hypersensitivity. To develop a plant whose nutrient uptake is improved with normal stomatal movement, we generated grafted plants (scion/rootstock : Col-0 (WT)/ost2-2D), and compared their growth and ionome profile with those of control plants (WT/WT) under nutrient-rich or nutrient-poor conditions. WT/ost2-2D shoots had larger fresh weight, dry weight, rosette diameter, leaf blade area, and contents of C, N, K, Ca, S, P, Mg, Na, Mn, B, Co, and Mo under nutrient-poor conditions compared with WT/WT shoots. The root fresh and dry weights and primary root length were greater in WT/ost2-2D roots than in WT/WT roots under both nutrient conditions. These results suggest that root-specific activation of PM H+-ATPase enhances root nutrient uptake, which accelerates the plant growth under nutrient-poor condition. Our study presents a novel approach to improving nutrient uptake efficiency in crops and vegetables; thus, it could contribute to the implementation of low-input sustainable agriculture.
Whole-transcriptomic profiling of human cerebral cortex tissues reveals microglia-associated molecular subtypes

Microglia is one of the major immune cell types in the human brain and plays pivotal roles in regulating inflammatory and immune response in healthy as well as disease states. By analyzing whole transcriptomic data derived from a large cohort of postmortem cortex tissues, we identified two distinct microglial subtypes within the population. The main difference between the two subtypes lies in the differential expression levels of the C1q complex components, Fc γ receptor (CD16) components and CD14. We validated our discovery in independent cohorts of brain autopsy tissues as well as in RNA-seq data generated from isolated microglia. Future investigations into the causes and physiological implications of these subtypes may shed more light on the homeostasis and regulation of the immune related processes in the brain.
Coordination of Pickpocket ion channel delivery and dendrite growth in Drosophila sensory neurons

Sensory neurons enable an organism to perceive external stimuli, which is essential for survival. The sensory capacity of a neuron depends on the elaboration of its dendritic arbor and the delivery of sensory ion channels to the dendritic membrane. However, it is not well understood how ion channels are trafficked to sensory dendrites and whether their delivery is coordinated with dendrite growth. We investigated the trafficking of the DEG/ENaC/ASIC ion channel Pickpocket (Ppk) in peripheral sensory neurons in fruit fly larvae. We used CRISPR-Cas9 genome engineering to tag endogenous Ppk1 and visualize it live, including monitoring Ppk1 membrane localization via a novel secreted split-GFP approach. Strikingly, Ppk1 is present throughout the membrane of actively growing dendrites, and Ppk1 density scales in proportion to the dendritic membrane, even when dynein-mediated transport to dendrites is disrupted. Our data suggest that Ppk1 is integral to the membrane of growing dendrites and implicate the recycling endosome GTPase Rab11 in the forward trafficking of Ppk1 to dendrites. Together, our results suggest that Ppk channel transport is coordinated with dendrite morphogenesis, thus ensuring proper ion channel levels and distribution in sensory dendrites. Author Summary Peripheral sensory neurons are essential for an organism to interact with its environment. Neurons are composed of signal-receiving dendrites and a signal-sending axon. Ion channels distributed throughout sensory dendrites transduce external stimuli into chemical signals, however the mechanisms that localize ion channels to sensory dendrites are not well understood. Both the composition of ion channels in the dendrites and the structure of a sensory neuron’s dendritic arbor are important for how it functions to sense external stimuli. Using live imaging and genomic engineering, we have discovered that the localization of a sensory ion channel, Pickpocket, in fruit fly sensory neurons is coordinated with growth of the dendritic arbor and that Pickpocket levels scale in proportion to dendrite length, even when transport to dendrites is disrupted. We also developed a novel genetically encoded approach to visualize the membrane expression of proteins in a living organism utilizing the split-GFP system. We found that both the secretory and endosomal networks mediate the forward trafficking of Pickpocket during neuronal morphogenesis, thus coordinating membrane growth with ion channel delivery. Our findings reveal that actively growing sensory dendrites are equipped with the ion channels needed for sensing external stimuli.
Exploring the Mosquito–Arbovirus Network: A Survey of Vector Competence Experiments

Arboviruses receive heightened research attention during major outbreaks, or when they cause unusual or severe clinical disease, but are otherwise under-characterized. Global change is also accelerating the emergence and spread of arboviral diseases, leading to time sensitive questions about potential interactions between viruses and novel vectors. Vector competence experiments help determine the susceptibility of certain arthropods to a given arbovirus, but these experiments are often conducted in real-time during outbreaks, rather than with preparedness in mind. We conducted a systematic review of reported mosquito-arbovirus competence experiments, screening 570 abstracts to arrive at 265 studies testing in vivo arboviral competence. We found that over 90% of potential mosquito-virus combinations are untested in experimental settings, and that entire regions and their corresponding vectors and viruses are undersampled. These knowledge gaps stymie outbreak response, and limit attempts to both build and validate predictive models of the vector-virus network.
Two different mental disc forms in the Tashan cave Garra: different species or ecotypes?

In the Tashan Cave barb Garra tashanensis, inhabiting a small cave in south west Iran, two mental disc forms were observed. To assess their phylogenetic relationships, disc-less and disc-bearing individuals were analyzed using mitochondrial cytochrome oxidase subunit I (COI) partial DNA sequences. Both mental disc forms nested within one clade with absolute bootstrap support (BS = 100), and the genetic distances between the disc-bearing and disc-less individuals (0.3-0.7%) was considerably lower than inter-species mtDNA sequence distances reported among members of the genus Garra. Hence, the observed mental disc variation was not inferred to be a taxonomic feature or a consequence of character displacement. Instead, it was inferred as a case of character release to diversify among ecological niches in the limited subterranean habitat, which should be clarified in follow-up ecological and population genetic studies in more detail.
Semi-supervised deep learning with graph neural network for cross-species regulatory sequence prediction

Genome-wide association studies have systematically identified thousands of single nucleotide polymorphisms (SNPs) associated with complex genetic diseases. However, the majority of those SNPs were found in non-coding genomic regions, preventing the understanding of the underlying causal mechanism. Predicting molecular processes based on the DNA sequence represents a promising approach to understand the role of those non-coding SNPs. Over the past years, deep learning was successfully applied to regulatory sequence prediction. Such method required DNA sequences associated with functional data for training. However, the human genome has a finite size which strongly limits the amount of DNA sequence with functional data available for training. Conversely, the amount of mammalian DNA sequences is exponentially increasing due to ongoing large sequencing projects, but without functional data in most cases. Here, we propose a semi-supervised learning approach based on graph neural network which allows to borrow information from homologous mammal sequences during training. Our approach can be plugged into any existing deep learning model and showed improvements in many different situations, including classification and regression, and for different types of functional data.
Larger cerebral cortex is genetically correlated with greater frontal area and dorsal thickness

Human cortical expansion has occurred non-uniformly across the brain. We assessed the genetic architecture of cortical global expansion and regionalization by comparing two sets of genome-wide association studies of 24 cortical regions with and without adjustment for global measures (i.e. total surface area, mean cortical thickness) using a genetically-informed parcellation in 32,488 adults. We found 393 and 756 significant loci with and without adjusting for globals, respectively, among which 8% and 45% loci were associated with more than one region. Results from analyses without adjustment for globals recounted loci associated with global measures. Genetic factors that contribute to total surface area of the cortex particularly expand anterior/frontal regions, whereas those contributing to thicker cortex predominantly increase dorsal/frontal-parietal thickness. Interactome-based analyses revealed significant overlap of global and regional genetic modules, enriched for neurodevelopmental and immune system pathways. Consideration of global measures is important in understanding the genetic variants underlying cortical morphology.
Non-aging despite high mutation rate – genomic insights into the evolution of Hydra

Hydra is a genus of freshwater polyps with remarkable regeneration abilities and a non-senescent phenotype under laboratory conditions. Thus, this animal is particularly interesting for aging research. Here, we gained insights into Hydra’s recent genetic evolution by genome sequencing of single cells and whole individuals. Despite its extreme longevity, Hydra does not show a lower somatic mutation rate than humans or mice. While we identify biological processes that have evolved under positive selection in animals kept in optimal laboratory conditions for decades, we found no signs of strong negative selection during this tiny evolutionary window. Interestingly, we observe the opposite pattern for the preceding evolution in the wild over a longer time period. Moreover, we found evidence that Hydra evolution in captivity was accompanied and potentially accelerated by loss of heterozygosity. Processes under positive selection in captive animals include pathways associated with Hydra’s simple nervous system, its nucleic acid metabolic process, cell migration, and hydrolase activity. Genes associated with organ regeneration, regulation of mRNA splicing, histone ubiquitination, and mitochondrial fusion were identified as highly conserved in the wild. Remarkably, several of the processes under strongest selection are closely related to those considered essential for the exapted, i. e. not brought about by natural selection, feature: Hydra’s non-aging.
Mechanosensor Piezo1 Mediates Bimodal Patterns of Intracellular Signaling

Piezo1 belongs to mechano-activatable cation channels serving as biological force sensors. However, the molecular events downstream of Piezo1 activation remain unclear. In this study, we used biosensors based on fluorescence resonance energy transfer (FRET) to investigate the dynamic modes of Piezo1-mediated signaling and revealed a bimodal pattern of Piezo1-induced intracellular calcium signaling. Laser-induced shockwaves (LIS) and its associated shear stress can mechanically activate Piezo1 to induce transient intracellular calcium (Ca[i]) elevation, accompanied by an increase in FAK activity. Interestingly, multiple pulses of shockwave stimulation caused a more sustained calcium increase and a decrease of FAK activity. Similarly, tuning the degree of Piezo1 activation by titrating either the dosage of Piezo1 ligand Yoda1 or the expression level of Piezo1, produced a similar bimodal pattern of FAK responses. Specifically, a low degree of Piezo1 activation (transient mode) leads to a transient Ca[i] response with FAK activation, whereas a high degree of Piezo1 activation (sustained mode) causes a sustained Ca[i] response with FAK suppression. Further investigation revealed that SHP2 serves as an intermediate regulator mediating this bimodal pattern in Piezo1 sensing and signaling. These results suggest that the degrees of Piezo1 activation induced by both mechanical LIS and chemical ligand stimulation may determine downstream signaling characteristics.
Held out wings RNA binding activity in the cytoplasm during early spermatogenesis

Held out wings (HOW) is an RNA-binding protein essential for spermatogenesis in Drosophila melanogaster. HOW is a signal transduction and activation of RNA (STAR) protein, regulating post-transcriptional gene expression. The characteristics of RNA-binding by the conserved short cytoplasmic isoform, HOW(S), are unknown. In vivo RIP-seq identified 121 novel transcripts bound by HOW(S) in germ stem cells and spermatogonia, many with signal transduction functions. (A/G/U)CUAAC motifs were enriched in 3’-UTRs and GCG(A/U)G in 5’-UTRs. HOW binds with high-affinity to sites containing CUAAC motifs from lola and hipk mRNAs. This study provides new insight into STAR protein-RNA interactions and functions in spermatogenesis.
Combined omics unravels the molecular mechanism of golden-leaf coloration in Koelreuteria paniculata ‘jinye’

Koelreuteria paniculata is widely distributed in Asia and introduced to Europe and North America. K. paniculata ‘jinye’ is a mutant variety used in landscaping that has a golden leaf color phenotype. Although similar leaf color variants occur in plants, little is known of the underlying mechanism. We performed physiological, anatomical, microRNA sequencing, transcriptomic, and metabolomic analyses of the golden leaf variation in the mutant. Compared with the original green cultivar, the golden leaf mutant exhibited 76.05% and 44.32% decreased chlorophyll a (Chl a) and chlorophyll b (Chl b) contents, respectively, and significantly increased carotenoid content. Analysis of leaf ultrastructure revealed an abnormal chloroplast morphology and fewer lamellae in the mutant. Fifty-nine differentially expressed genes (DEGs), forty transcription factors (TFs) and forty-nine differentially expressed miRNAs (DEmiRs) involved in pigment metabolism, chloroplast development, and photosynthesis were identified. The GLK and petC genes were downregulated and are involved in chloroplast development and chlorophyll synthesis, respectively. The upregulated PSY and PDS genes, and the downregulated NCED gene promote carotenoid accumulation. A variety of chalcones and flavonols were upregulated in the mutant. Consequently, the carotenoid to chlorophyll ratio increased by more than 75%, and the accumulation of chalcones and flavonols was responsible for the golden leaf phenotype of the mutant K. paniculata.
ResPAN: a powerful batch correction model for scRNA-seq data through residual adversarial networks

Motivation With the advancement of technology, we can generate and access large-scale, high dimensional and diverse genomics data, especially through single-cell RNA sequencing (scRNA-seq). However, integrative downstream analysis from multiple scRNA-seq datasets remains challenging due to batch effects. Results In this paper, we propose a light-structured deep learning framework called ResPAN for scRNA-seq data integration. ResPAN is based on Wasserstein Generative Adversarial Network (WGAN) combined with random walk mutual nearest neighbor pairing and fully skip-connected autoencoders to reduce the differences among batches. We also discuss the limitations of existing methods and demonstrate the advantages of our model over seven other methods through extensive benchmarking studies on both simulated data under various scenarios and real datasets across different scales. Our model achieves leading performance on both batch correction and biological information conservation and maintains scalable to datasets with over half a million cells. Availability An open-source implementation of ResPAN and scripts to reproduce the results can be downloaded from: https://github.com/AprilYuge/ResPAN. Contact hongyu.zhao@yale.edu
MCell4 with BioNetGen: A Monte Carlo Simulator of Rule-Based Reaction-Diffusion Systems with Python Interface

Biochemical signaling pathways in living cells are often highly organized into spatially segregated volumes, membranes, scaffolds, subcellular compartments, and organelles comprising small numbers of interacting molecules. At this level of granularity stochastic behavior dominates, well-mixed continuum approximations based on concentrations break down and a particle-based approach is more accurate and more efficient. We describe and validate a new version of the open-source MCell simulation program (MCell4), which supports generalized 3D Monte Carlo modeling of diffusion and chemical reaction of discrete molecules and macromolecular complexes in solution, on surfaces representing membranes, and combinations thereof. The main improvements in MCell4 compared to the previous versions, MCell3 and MCell3-R, include a Python interface and native BioNetGen reaction language (BNGL) support. MCell4’s Python interface opens up completely new possibilities for interfacing with external simulators to allow creation of sophisticated event-driven multiscale/multiphysics simulations. The native BNGL support, implemented through a new open-source library libBNG (also introduced in this paper), provides the capability to run a given BNGL model spatially resolved in MCell4 and, with appropriate simplifying assumptions, also in the BioNetGen simulation environment, greatly accelerating and simplifying model validation and comparison.
Two therapeutic CRISPR/Cas9 gene editing approaches revert FTD/ALS cellular pathology caused by a C9orf72 repeat expansion mutation in patient derived cells

CRISPR gene editing holds promise to cure or arrest genetic disease, if we can find and implement curative edits reliably, safely and effectively. Expansion of a hexanucleotide repeat in C9orf72 is the leading known genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). We evaluated three approaches to editing the mutant C9orf72 gene for their ability to correct pathology in neurons derived from patient iPSCs: excision of the repeat region, excision of the mutant allele, and excision of regulatory region exon 1A. All three approaches normalized RNA abnormalities and TDP-43 pathology, but only repeat excision and mutant allele excision completely eliminated pathologic dipeptide repeats. Our work sheds light on the complex regulation of the C9orf72 gene and suggests that because of sense and anti-sense transcription, silencing a single regulatory region may not reverse all pathology. Our work also provides a roadmap for evaluating CRISPR gene correction using patient iPSCs.
Functional gene analysis and cultivation experiments predict the degradation of diverse extracellular polysaccharides by ubiquitous taxa in pustular microbial mats from Shark Bay, Western Australia

Microbial exopolymeric substances (EPSs) form the organic, polysaccharide-rich matrix of marine microbial mats and can mediate the binding and precipitation of carbonate minerals therein. Here, we investigate the molecular ecology of carbohydrate degradation and production in pustular mats from Shark Bay, Western Australia, by analyzing 84 metagenome-assembled genomes (MAGs) and the composition of microbial communities enriched from a pustular mat on various polysaccharide substrates. The annotation of 4000 genes from hundreds of carbohydrate-active enzyme (CAZyme) families in the MAGs and mapping of polysaccharide-degrading CAZymes to their predicted substrates identify trends in the distribution and localization of degradation-associated CAZymes across different bacterial phyla. The compositions of microbial communities enriched on a range of polysaccharides inoculated with pustular mat material support the predicted trends. The combined metagenomic and experimental analyses reveal a widespread potential for EPS degradation among MAGs from Shark Bay pustular mats and suggest distinct roles for some phyla that are reported at high abundances in mats. Specifically, Bacteroidetes are likely to be primary degraders of polysaccharide EPSs, alongside Planctomycetes and a small subset of Alphaproteobacteria and Gammaproteobacteria. Planctomycetes, some Bacteroidetes, Verrucomicrobia, Myxococcota and Anaerolineae are also predicted to favor degradation of sulfated substrates, which are present in the EPS matrix of pustular mats. Large sets of functionally varied CAZymes without signal peptides tagging them for export implicate Anaerolineae and Verrucomicrobia in degrading the downstream products of primary EPS degradation. Importance Modern marine microbial mats are rich in exopolymeric substances (EPSs) — complex, high molecular weight polymers secreted by bacteria — that mediate the formation of carbonate minerals and the preservation of microbial textures in mats. However, the organisms involved in EPS cycling in these mats have not been identified and the links between EPS degradation, carbonate precipitation, and microbial ecology in mats remain poorly understood. We define distinct roles in EPS cycling for many major microbial taxa that are both ubiquitous and abundant in pustular microbial mats from Shark Bay, Australia. The large genomic potential of these microbes for the modification and degradation of diverse extracellular organic polymers provides a blueprint for future studies aimed at quantifying and verifying the specific contributions of these microbes to EPS degradation, carbon cycling and carbonate precipitation.