Synthetic droplets mimicking bio-soft matter droplets formed via liquid-liquid phase separation (LLPS) in living cells have recently been employed in nanobiotechnology for artificial cells, molecular ...robotics, molecular computing, etc. Temporally controlling the dynamics of synthetic droplets is essential for developing such bio-inspired systems because living systems maintain their functions based on the temporally controlled dynamics of biomolecular reactions and assemblies. This paper reports the temporal control of DNA-based LLPS droplets (DNA droplets). We demonstrate the timing-controlled division of DNA droplets via time-delayed division triggers regulated by chemical reactions. Controlling the release order of multiple division triggers results in order control of the multistep droplet division, i.e., pathway-controlled division in a reaction landscape. Finally, we apply the timing-controlled division into a molecular computing element to compare microRNA concentrations. We believe that temporal control of DNA droplets will promote the design of dynamic artificial cells/molecular robots and sophisticated biomedical applications.Controlling the dynamics of synthetic liquid-liquid phase separation droplets is essential for various bioinspired systems. Here, authors demonstrate temporally-controlled multi-step division of DNA-based liquid droplets, and develop a molecular computation system to compare miRNA concentrations.
Background & Aims
In about 20% of children with cholestasis and normal or low serum gamma‐glutamyltransferase (GGT) activity, no aetiology is identified. We sought new genes implicated in paediatric ...hepatobiliary disease.
Methods
We conducted whole‐exome sequencing in 69 children evaluated at our centre from 2011 to 2018 who had low‐GGT cholestasis and in whom homozygous/compound heterozygous predictedly pathogenic variants (PPVs) in ATP8B1, ABCB11, NR1H4, MYO5B or TJP2 were not found. Clinical records and findings on light microscopy and transmission electron microscopy of liver biopsy materials were reviewed.
Results
In seven patients from seven unrelated families, biallelic PPVs (10 in total) were found in USP53, recently associated with intrahepatic cholestasis. Seven variants were classified as pathogenic: one canonical splicing, c.569 + 2T > C, and six nonsense or frameshifting: c.169C > T (p.Arg57Ter), c.581delA (p.Arg195GlufsTer38), c.831_832insAG (p.Val279GlufsTer16), c.1012C > T (p.Arg338Ter), c.1426C > T (p.Arg476Ter) and c.1558C > T (p.Arg520Ter). Three were likely pathogenic: c.297G > T (p.Arg99Ser), c.395A > G (p.His132Arg) and c.878G > T (p.Gly293Val). In all patients, jaundice began at age <7 months. Cholestasis was transient, with documented resolution of hyperbilirubinaemia in all (oldest patient now aged 5 years) except one, who was lost to follow‐up. Light microscopy identified intralobular cholestasis, giant‐cell change of hepatocytes and perisinusoidal‐perihepatocytic and portal‐tract fibrosis. Ultrastructural study revealed elongated hepatocyte‐hepatocyte tight junctions. One patient was deaf.
Conclusion
USP53 interacts with the tight junction constituent TJP2. TJP2 mutation can cause low‐GGT intrahepatic cholestasis, with elongated hepatocyte‐hepatocyte tight junctions, as well as deafness. Our findings extend a preliminary report of USP53 disease and indicate that USP53 mutation may generate a partial phenocopy of TJP2 disease.
Long–non‐coding RNAs (lncRNA) AWPPH promotes the progression of liver and bladder cancer, indicating its oncogenic role. The current study aimed to explore the involvement of AWPPH in triple‐negative ...breast cancer (TNBC). In the current study, we found that plasma levels of lncRNA AWPPH and microRNA‐21 (miRNA‐21) were upregulated in patients with TNBC than in healthy controls, and the upregulation of plasma lncRNA AWPPH and miRNA‐21 distinguished early‐stage patients with TNBC from healthy controls. Plasma levels of lncRNA AWPPH and miRNA‐21 were significantly and positively correlated in both patients with TNBC and healthy controls. LncRNA AWPPH and miRNA‐21 overexpression led to promoted cancer cells proliferation and improved cancer cell viability under carboplatin treatment, while lncRNA AWPPH small interfering RNA (siRNA) silencing played an opposite role. In addition, miRNA‐21 overexpression attenuated the effects of lncRNA AWPPH siRNA silencing on of cancer cell behaviors. LncRNA AWPPH overexpression led to upregulated miRNA‐21 in TNBC cells, while miRNA‐21 overexpression also led to significantly upregulated lncRNA AWPPH expression. Therefore, lncRNA AWPPH and miRNA‐21 may regulate cancer cell proliferation and chemosensitivity in TNBC by interacting with each other.
Long–non‐coding RNA (lncRNA) AWPPH and microRNA‐21 (miRNA‐21) were both upregulated in triple‐negative breast cancer (TNBC). LncRNA AWPPH and miRNA‐21 may regulate cancer cell proliferation and chemosensitivity in TNBC by interacting with each other.
Phase‐separated biomolecular droplets are formed in cells to regulate various biological processes. This phenomenon can be applied to constructing self‐assembled dynamic molecular systems such as ...artificial cells and molecular robots. Recently, programmable phase‐separated droplets called DNA droplets have been reported as a possible method to construct such dynamic molecular systems. This study reports a computational DNA droplet that can recognize a specific combination of tumor biomarker microRNAs (miRNAs) as molecular inputs and output a DNA logic computing result by physical DNA droplet phase separation. A mixed DNA droplet consisting of three DNA nanostructures with orthogonal sticky‐end sequences and two linker DNAs to cross‐bridge the orthogonal DNA nanostructures is proposed. By the hybridization of miRNAs with the linkers, the cross‐bridging ability is lost, causing the phase‐separation of the mixed DNA droplet into three DNA droplets, resulting in executing a miRNA pattern recognition described by a logical expression ((miRNA‐1 ∧ miRNA‐2) ∧ (miRNA‐3 ∧ ¬miRNA‐4)). This experimentally demonstrates that the computational DNA droplets recognize the above specific pattern of chemically synthesized miRNA sequences as a model experiment. In the future, this method will provide potential applications such as diagnosis and therapy with integration to biomolecular robots and artificial cells.
Computational DNA droplets recognizing a specific combination of tumor biomarker microRNAs (miRNAs) are demonstrated. The computational DNA droplet can execute DNA logic operation ((miRNA‐1 ∧ miRNA‐2) ∧ (miRNA‐3 ∧ ¬miRNA‐4)). A uniformly mixed DNA droplet can output the computational results by nucleic acid‐triggered liquid–liquid phase separation of DNA droplets into three droplets.
The recently emerged ferromagnetic two-dimensional (2D) materials provide unique platforms for compact spintronic devices down to the atomic-thin regime; however, the prospect is hindered by the ...limited number of ferromagnetic 2D materials discovered with limited choices of magnetic properties. If 2D antiferromagnetism could be converted to 2D ferromagnetism, the range of 2D magnets and their potential applications would be significantly broadened. Here, we discovered emergent ferromagnetism by interfacing non-magnetic WS
layers with the antiferromagnetic FePS
. The WS
exhibits an order of magnitude enhanced Zeeman effect with a saturated interfacial exchange field ~38 Tesla. Given the pristine FePS
is an intralayer antiferromagnet, the prominent interfacial exchange field suggests the formation of ferromagnetic FePS
at interface. Furthermore, the enhanced Zeeman effect in WS
is found to exhibit a strong WS
-thickness dependence, highlighting the layer-tailorable interfacial exchange coupling in WS
-FePS
heterostructures, which is potentially attributed to the thickness-dependent interfacial hybridization.
The shift current (SHC) has been accepted as the primary mechanism of the bulk photovoltaic effect (BPVE) in ferroelectrics, which is much different from the typical p-n junction-based photovoltaic ...mechanism in heterogeneous materials. In the present work, we use first-principles calculations to investigate the SHC response in the ferroelectric semiconductor GeTe, which is found possess a large SHC response due to its intrinsic narrow band gap and high covalency. We explore the changes of SHC response induced by phonon vibrations, and analytically fit current versus vibrational amplitude to reveal the quantitative relationships between vibrations and the SHC response. Furthermore, we demonstrate the temperature dependence of the SHC response by averaging the phonon vibration influence in the Brillouin zone. Our investigation provides an explicit experimental prediction about the temperature dependence of BPVE and can be extended to other classes of noncentrosymmetric materials.
Effective and safe delivery of the CRISPR/Cas9 gene-editing elements remains a challenge. Here we report the development of PEGylated nanoparticles (named P-HNPs) based on the cationic α-helical ...polypeptide poly(γ-4-((2-(piperidin-1-yl)ethyl)aminomethyl)benzyl-L-glutamate) for the delivery of Cas9 expression plasmid and sgRNA to various cell types and gene-editing scenarios. The cell-penetrating α-helical polypeptide enhanced cellular uptake and promoted escape of pCas9 and/or sgRNA from the endosome and transport into the nucleus. The colloidally stable P-HNPs achieved a Cas9 transfection efficiency up to 60% and sgRNA uptake efficiency of 67.4%, representing an improvement over existing polycation-based gene delivery systems. After performing single or multiplex gene editing with an efficiency up to 47.3% in vitro, we demonstrated that P-HNPs delivering Cas9 plasmid/sgRNA targeting the polo-like kinase 1 (Plk1) gene achieved 35% gene deletion in HeLa tumor tissue to reduce the Plk1 protein level by 66.7%, thereby suppressing the tumor growth by >71% and prolonging the animal survival rate to 60% within 60 days. Capable of delivering Cas9 plasmids to various cell types to achieve multiplex gene knock-out, gene knock-in, and gene activation in vitro and in vivo, the P-HNP system offers a versatile gene-editing platform for biological research and therapeutic applications.
Enhancer RNA (eRNA) is a type of noncoding RNA transcribed from the enhancer. Although critical roles of eRNA in gene transcription control have been increasingly realized, the systemic landscape and ...potential function of eRNAs in cancer remains largely unexplored. Here, we report the integration of multi-omics and pharmacogenomics data across large-scale patient samples and cancer cell lines. We observe a cancer-/lineage-specificity of eRNAs, which may be largely driven by tissue-specific TFs. eRNAs are involved in multiple cancer signaling pathways through putatively regulating their target genes, including clinically actionable genes and immune checkpoints. They may also affect drug response by within-pathway or cross-pathway means. We characterize the oncogenic potential and therapeutic liability of one eRNA, NET1e, supporting the clinical feasibility of eRNA-targeted therapy. We identify a panel of clinically relevant eRNAs and developed a user-friendly data portal. Our study reveals the transcriptional landscape and clinical utility of eRNAs in cancer.
Allylation and propargylation are two powerful synthetic strategies for making new substances that have been of significant importance in chemistry, medicine, and material fields. Conventional ...tactics employ various preformed allylation and propargylation reagents. In this study, a conceptually novel copper-catalyzed and B2pin2-mediated Umpolung reactivity of propargylic carbonates has been achieved for the first time, realizing both allylation and propargylation of aldehydes and ketones without additional reductants. Three types of allylation products and one type of propargylation product are generated efficiently, and all allylation products are formed with syn-configurations predominantly. The choice of ligands plays a vital role in modulating the Umpolung modes. The synthetic applications have been demonstrated in a myriad of further transformations including natural product synthesis, and systematic mechanistic studies have been conducted to reveal detailed insights into the Umpolung processes.
Tremendous progress has been made in the field of toxicology leading to the advance of developmental toxicity assessment. Conventional animal models and in vitro two-dimensional models cannot ...accurately describe toxic effects and predict actual in vivo responses due to obvious inter-species differences between humans and animals, as well as the lack of a physiologically relevant tissue microenvironment. Human embryonic stem cell (hESC)- and induced pluripotent stem cell (iPSC)-derived three-dimensional organoids are ideal complex and multicellular organotypic models, which are indispensable in recapitulating morphogenesis, cellular interactions, and molecular processes of early human organ development. Recently, human organoids have been used for drug discovery, chemical toxicity and safety in vitro assessment. This review discusses the recent advances in the use of human organoid models, (i.e., brain, retinal, cardiac, liver, kidney, lung, and intestinal organoid models) for developmental toxicity and teratogenicity assessment of distinct tissues/organs following exposure to pharmaceutical compounds, heavy metals, persistent organic pollutants, nanomaterials, and ambient air pollutants. Combining next-generation organoid models with innovative engineering technologies generates novel and powerful tools for developmental toxicity and teratogenicity assessment, and the rapid progress in this field is expected to continue.
•Advances in stem cell techniques for toxicity assessment.•Human organoids can decipher the embryonic development and morphogenesis.•Organoid technologies for developmental toxicology assessment.•Advanced next-generation human organoids toward next-level toxicity testing.
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