CRISPR-mediated transcriptional activation (CRISPRa) is a powerful technology for inducing gene expression from endogenous loci with exciting applications in high throughput gain-of-function genomic ...screens and the engineering of cell-based models. However, current strategies for generating potent, stable, CRISPRa-competent cell lines present limitations for the broad utility of this approach. Here, we provide a high-efficiency, self-selecting CRISPRa enrichment strategy, which combined with piggyBac transposon technology enables rapid production of CRISPRa-ready cell populations compatible with a variety of downstream assays. We complement this with an optimized guide RNA scaffold that significantly enhances CRISPRa functionality. Finally, we describe a synthetic guide RNA tool set that enables transient, population-wide gene activation when used with the self-selecting CRISPRa system. Taken together, this versatile platform greatly enhances the potential for CRISPRa across a wide variety of cellular contexts.
Vascular endothelial growth factor inhibition has become a cornerstone of cancer therapy. Significant adverse effect is associated with these therapies, with hypertension among the most prevalent. We ...aim to provide a current review of the mechanisms by which these therapies induce hypertension, the risk factors and predictors of the development of hypertension, and management strategies to minimize the risk of cardiovascular complications in patients receiving vascular endothelial growth factor inhibition as part of cancer treatment.
Fish skin contains a mucosal microbiome for the largest and oldest group of vertebrates, a location ideal for microbial community ecology and practical applications in agriculture and veterinary ...medicine. These selective microbiomes are dominated by
Proteobacteria
, with compositions different from the surrounding water. Core taxa are a small percentage of those present and are currently functionally uncharacterized. Methods for skin sampling, DNA extraction and amplification, and sequence data processing are highly varied across the field, and reanalysis of recent studies using a consistent pipeline revealed that some conclusions did change in statistical significance. Further, the 16S gene sequencing approaches lack quantitation of microbes and copy number adjustment. Thus, consistency in the field is a serious limitation in comparing across studies. The most significant area for future study, requiring metagenomic and metabolomics data, is the biochemical pathways and functions within the microbiome community, the interactions between members, and the resulting effects on fish host health being linked to specific nutrients and microbial species. Genes linked to skin colonization, such as those for attachment or mucin degradation, need to be uncovered and explored. Skin immunity factors need to be directly linked to microbiome composition and individual taxa. The basic foundation has been laid, and many exciting future discoveries remain.
Advanced high-grade serous ovarian cancer (HGSC) is an aggressive disease that accounts for 70% of all ovarian cancer deaths. Nevertheless, 15% of patients diagnosed with advanced HGSC survive more ...than 10 years. The elucidation of predictive markers of these long-term survivors (LTS) could help identify therapeutic targets for the disease, and thus improve patient survival rates. To investigate the stromal heterogeneity of the tumor microenvironment (TME) in ovarian cancer, we used spatial transcriptomics to generate spatially resolved transcript profiles in treatment-naïve advanced HGSC from LTS and short-term survivors (STS) and determined the association between cancer-associated fibroblasts (CAF) heterogeneity and survival in patients with advanced HGSC. Spatial transcriptomics and single-cell RNA-sequencing data were integrated to distinguish tumor and stroma regions, and a computational method was developed to investigate spatially resolved ligand-receptor interactions between various tumor and CAF subtypes in the TME. A specific subtype of CAFs and its spatial location relative to a particular ovarian cancer cell subtype in the TME correlated with long-term survival in patients with advanced HGSC. Also, increased APOE-LRP5 cross-talk occurred at the stroma-tumor interface in tumor tissues from STS compared with LTS. These findings were validated using multiplex IHC. Overall, this spatial transcriptomics analysis revealed spatially resolved CAF-tumor cross-talk signaling networks in the ovarian TME that are associated with long-term survival of patients with HGSC. Further studies to confirm whether such cross-talk plays a role in modulating the malignant phenotype of HGSC and could serve as a predictive biomarker of patient survival are warranted.
Generation of spatially resolved gene expression patterns in tumors from patients with ovarian cancer surviving more than 10 years allows the identification of novel predictive biomarkers and therapeutic targets for better patient management. See related commentary by Kelliher and Lengyel, p. 1383.
Context.
Pulsars show a steady decrease in their rotational frequency, occasionally interrupted by sudden spin-ups called glitches, whose physical origin is still a mystery. One suggested explanation ...for at least the small glitches are starquakes, that is, failures of the solid neutron star crust, in which the progressive reduction in the centrifugal force deforms the star, stressing the solid until it breaks. This produces a spin-up, dissipating energy inside the star.
Aims.
We explore this suggestion by analyzing a mostly analytical model in order to understand the possible consequences of starquakes, particularly whether they can explain at least the small glitches.
Methods.
We analyze the deformations and strains produced by the decreasing centrifugal force, modeling the neutron star with a fluid core and a solid crust, each with uniform density and with the core possibly denser than the crust, as a simple approximation to the strong density gradient present in real neutron stars.
Results.
The deformation of a star with very different densities in the core and crust is qualitatively different from the previously studied case of equal densities. The former more closely resembles the behavior of a fluid star, in which the core-crust interface is a surface of constant gravitational plus centrifugal potential.
Conclusions.
Regardless of the uncertain breaking strain, the glitch activity in this model is several orders of magnitude smaller than observed, even if only small glitches are considered. For a large breaking strain, suggested by simulations, glitches due to starquakes could be roughly of the correct size but much less frequent than observed glitches. The energy released in each such glitch is much larger than in the standard model of angular momentum transfer from a faster rotating superfluid in the inner crust. On the other hand, we cannot rule out that the heating produced by small starquakes could trigger glitches by allowing neutron superfluid vortices to move. We also confirm that stresses in the neutron star crust can in principle support an ellipticity much larger than some observational upper limits from pulsar timing and continuous gravitational wave searches.
FU Orionis stars (FUors) are eruptive pre-main sequence objects thought to represent quasi-periodic or recurring stages of enhanced accretion during the low-mass star-forming process. We compare our ...models with those obtained by Robitaille et al. for Taurus class II and I sources in quiescence periods by calculating the cumulative distribution of the different parameters. FUors have more massive disks: we find that ~80% of the disks in FUors are more massive than any Taurus class II and I sources in the sample. Median values for the disk mass accretion rates are ~10 super(-7) M sub(e) yr super(-1) versus ~10 super(-5) M sub(odot) yr super(-1) for standard young stellar objects (YSOs) and FUors, respectively. Our results support the current scenario in which changes experimented by the circumstellar disk explain the observed properties of these stars. However, the increase in the disk mass accretion rate is smaller than theoretically predicted, although in good agreement with previous determinations.
Two heterometallic photocatalysts were designed and probed for water reduction. Both (bpy)2RuIINiII(L1)(ClO4)2 (1) and (bpy)2RuIINiII(L2)2RuII(bpy)2(ClO4)2 (2) can generate the low‐valent precursor ...involved in hydride formation prior to dihydrogen generation. However, while the bimetallic RuIINiII (1) requires the presence of an external photosensitizer to trigger catalytic activity, the trimetallic RuIINiIIRuII (2) displays significant coupling between the catalytic and light‐harvesting units to promote intramolecular multielectron transfer and perform photocatalysis at the Ni center. A concerted experimental and theoretical effort proposes mechanisms to explain why 1 is unable to achieve self‐supported catalysis, while 2 is fully photocatalytic.
Two new heterometallic photocatalysts were probed for water reduction. While the bimetallic RuIINiII requires an external photosensitizer to be catalytic, the trinuclear heterometallic RuIINiIIRuII produces dihydrogen when irradiated with blue light. Mechanisms are proposed to account for these observations.
Childhood obesity linked to metabolic alterations, tend to appear simultaneously with altered adipocytokines, suggesting a role in pathogenetic development. Low circulating level of total and high ...molecular weight (HMW) adiponectin have been associated with components of the metabolic syndrome (MetS) and could represent an independent risk factor with potential use as a biomarker. To examine the prevalence of MetS in Mexican school children and to investigate the association of total and HMW adiponectin levels with biochemical parameters related to MetS.
The study included a population of boys and girls, from 8 to 11 years old. Anthropometric and biochemical parameters were evaluated according to weight and MetS status. A correlation analysis was fitted to establish an association between adiponectin concentrations and metabolic indicators.
One-hundred and fifty five children participated (59.4% females) from 8-11 years of age. The prevalence of MetS was of 10.3%. Impaired biochemical parameters, including total and HMW adiponectin, were associated with obesity. The adiponectin level was significantly lower in MetS than in non-MetS subjects (4.5 vs. 5.4 μg/mL). Total- but not HMW adiponectin concentration was negatively correlated with blood pressure, fasting insulin, fasting blood sugar and Homeostatic Model Assessment for Insulin Resistance.
In young children, the total adiponectin level is associated with impaired biochemical parameters of carbohydrate metabolism and could be an excellent early predictor of metabolic complications.
Inactivating TP53 mutations leads to a loss of function of p53, but can also often result in oncogenic gain-of-function (GOF) of mutant p53 (mutp53) proteins which promotes tumor development and ...progression. The GOF activities of TP53 mutations are well documented, but the mechanisms involved remain poorly understood. Here, we study the mutp53 interactome and find that by targeting minichromosome maintenance complex components (MCMs), GOF mutp53 predisposes cells to replication stress and chromosomal instability (CIN), leading to a tumor cell-autonomous and cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING)-dependent cytosolic DNA response that activates downstream non-canonical nuclear factor kappa light chain enhancer of activated B cell (NC-NF-κB) signaling. Consequently, GOF mutp53-MCMs-CIN-cytosolic DNA-cGAS-STING-NC-NF-κB signaling promotes tumor cell metastasis and an immunosuppressive tumor microenvironment through antagonizing interferon signaling and regulating genes associated with pro-tumorigenic inflammation. Our findings have important implications for understanding not only the GOF activities of TP53 mutations but also the genome-guardian role of p53 and its inactivation during tumor development and progression.
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