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1.
Small Methods ; : e2400084, 2024 May 13.
Article in English | MEDLINE | ID: mdl-38738733

ABSTRACT

Doping plays a crucial role in modulating and enhancing the performance of organic semiconductor (OSC) devices. In this study, the critical role of dopants is underscored in shaping the morphology and structure of OSC films, which in turn profoundly influences their properties. Two dopants, trityl tetrakis(pentafluorophenyl) (TrTPFB) and N,N-dimethylanilinium tetrakis(pentafluorophenyl)borate (DMA-TPFB), are examined for their doping effects on poly(3-hexylthiophene) (P3HT) and PBBT-2T host OSCs. It is found that although TrTPFB exhibits higher doping efficiency, OSCs doped with DMA-TPFB achieve comparable or even enhanced electrical conductivity. Indeed, the electrical conductivity of DMA-TPFB-doped P3HT reaches over 67 S cm-1, which is a record-high value for mixed-solution-doped P3HT. This can be attributed to DMA-TPFB inducing a higher degree of crystallinity and reduced structural disorder. Moreover, the beneficial impact of DMA-TPFB on the OSC films' morphology and structure results in superior thermoelectric performance in the doped OSCs. These findings highlight the significance of dopant-induced morphological and structural considerations in enhancing the film characteristics of OSCs, opening up a new avenue for optimization of dopant performance.

2.
Small ; 20(1): e2304626, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37641178

ABSTRACT

Electronics have greatly promoted the development of modern society and the exploration of new semiconducting materials with low cost and high mobility continues to attract interest in the advance of next-generation electronic devices. Among emerging semiconductors, the metal-halide perovskite, especially the nontoxic tin (Sn)-based candidates, has recently made breakthroughs in the field of diverse electronic devices due to its excellent charge transport properties and cost-effective large-area deposition capability at low temperatures. To enable a more comprehensive understanding of this emerging research field and promote the development of new-generation perovskite electronics, this review aims to provide an in-depth understanding with the discussion of unique physical properties of Sn-based perovskites and the summarization of recent research progress of Sn-based perovskite field-effect transistors (FETs) and diverse electronic devices. The unique character of negligible ion migration is also discussed, which is fundamentally different from the lead-based counterparts and provides a great prerequisite for device application. The following section highlights the potential broad applications of Sn-perovskite FETs as a competitive and feasible technology. Finally, an outlook and remaining challenges are given to advance the progression of Sn-based perovskite FETs, especially on the origin and solution of stability problems toward high-performance Sn-based perovskite electronics.

3.
Proc Natl Acad Sci U S A ; 119(45): e2211142119, 2022 Nov 08.
Article in English | MEDLINE | ID: mdl-36322771

ABSTRACT

Ultradian rhythms in metabolism and physiology have been described previously in mammals. However, the underlying mechanisms for these rhythms are still elusive. Here, we report the discovery of temperature-sensitive ultradian rhythms in mammalian fibroblasts that are independent of both the cell cycle and the circadian clock. The period in each culture is stable over time but varies in different cultures (ranging from 3 to 24 h). We show that transient, single-cell metabolic pulses are synchronized into stable ultradian rhythms across contacting cells in culture by gap junction-mediated coupling. Coordinated rhythms are also apparent for other metabolic and physiological measures, including plasma membrane potential (Δψp), intracellular glutamine, α-ketoglutarate, intracellular adenosine triphosphate (ATP), cytosolic pH, and intracellular calcium. Moreover, these ultradian rhythms require extracellular glutamine, several different ion channels, and the suppression of mitochondrial ATP synthase by α-ketoglutarate, which provides a key feedback mechanism. We hypothesize that cellular coupling and metabolic feedback can be used by cells to balance energy demands for survival.


Subject(s)
Circadian Clocks , Ultradian Rhythm , Animals , Ketoglutaric Acids , Glutamine , Cell Cycle , Circadian Rhythm/physiology , Mammals
4.
J Colloid Interface Sci ; 606(Pt 1): 800-807, 2022 Jan 15.
Article in English | MEDLINE | ID: mdl-34419819

ABSTRACT

All-inorganic perovskite materials (Typically: CsPbI2Br) have attracted enormous attention due to their illustrious thermal stability and appropriate bandgap, and their use in perovskite solar cells (PSCs) has been extensively investigated. However, the inevitable defects of the perovskite layer, energy level mismatch between perovskite and carbon electrodes, and the phase instability of CsPbI2Br limit the power conversion efficiency (PCE) and stability of carbon-based CsPbI2Br PSCs. Herein, we demonstrate a simple and effective strategy for regulating energy level, inhibiting carrier recombination, and delaying the degradation of perovskite by modifying the surface of CsPbI2Br with a new type of 2D perovskite Cs2PtI6. The carbon-based CsPbI2Br PSCs achieve a higher PCE (13.69 %) than the control device (11.10 %). The excellent matching of the energy level and suppression of charge carrier recombination should be responsible for the improvement in efficiency. Furthermore, the excellent hydrophobic performance of Cs2PtI6 enhances the moisture resistance of the device. This study provides a potential strategy for improving the performance and stability of all-inorganic CsPbI2Br PSCs.

5.
ChemSusChem ; 14(21): 4776-4782, 2021 Nov 04.
Article in English | MEDLINE | ID: mdl-34435455

ABSTRACT

Power conversion efficiencies (PCEs) of up to 25.5 % have been reported for perovskite solar cells (PSCs). Thus, they have shown great potential for commercial applications. Therefore, simplifying technological process and reducing production costs have been a widespread concern among scientific and industrial communities. In this study, PSCs are prepared with the simplest device architecture (FTO/MAPbI3 /carbon). A high-quality perovskite film with few interface defects and good carrier transport is obtained by tuning the p-n properties, matching energy levels, and enhancing carrier collection and transport. A PCE of 12.01 % is achieved, which is the best reported to date for this device structure. The device also shows excellent long-term stability, owing to the elimination of charge transport layers and the usage of hydrophobic materials. This study provides a new approach to reduce production costs and simplify production of PSCs.

6.
Nanoscale Adv ; 3(7): 1910-1916, 2021 Apr 06.
Article in English | MEDLINE | ID: mdl-36133092

ABSTRACT

In the family of inorganic perovskite solar cells (PSCs), CsPbBr3 has attracted widespread attention due to its excellent stability under high humidity and high temperature conditions. However, power conversion efficiency (PCE) improvement of CsPbBr3-based PSCs is markedly limited by the large optical absorption loss coming from the wide band gap and serious charge recombination at interfaces and/or within the perovskite film. In this work, using density functional theory calculations, we systemically studied the electronic properties of niobium (Nb)-doped CsPbBr3 with different concentration ratios. As a result, it is found that doped CsPbBr3 compounds are metallic at high Nb doping concentration but semiconducting at low Nb doping concentration. The calculated electronic density of states shows that the conduction band is predominantly constructed of doped Nb. These characteristics make them very suitable for solar cell and energy storage applications.

7.
Biochemistry ; 59(45): 4344-4352, 2020 11 17.
Article in English | MEDLINE | ID: mdl-33147009

ABSTRACT

Gemcitabine (dFdC), a modified deoxycytidine (dC) widely used in tumor treatment, is a prodrug that is phosphorylated to generate mono-, di-, and triphosphates. The triphosphate (dFdCTP) is incorporated into DNA to terminate DNA synthesis in cancer. Some incorporated dFdC nucleotides can be partially removed by the 3'-5' exonuclease activity, namely its editing function, and the others escape the editing. However, whether there is an active mechanism for dFdC to escape the editing remains unclear. We have first discovered that unlike dFdC, its mono-, di-, and triphosphates can inhibit the 3'-5' exonuclease of DNA polymerase I, suppress editing, and allow the active escaping mechanism, whereas its polymerase activity is not remarkably affected. As such, these phosphates can prevent the removal of the incorporated dFdC residue, thereby actively blocking DNA extension and synthesis. The inhibition efficiency of these phosphates follows the increased order of the mono-, di-, and triphosphates of gemcitabine (dFdC < dFdCMP < dFdCDP < dFdCTP). In addition, after the deletion of the 3'-5' exonuclease of cellular DNA polymerase I, the Escherichia coli mutant is more sensitive to dFdCTP than is wild-type E. coli. Our new discovery of the ability of these dFdC phosphates to inhibit exonuclease activity suggests a novel anticancer mechanism of gemcitabine and its phosphate derivatives.


Subject(s)
DNA/chemistry , Deoxycytidine/analogs & derivatives , Exonucleases/antagonists & inhibitors , Phosphates/chemistry , Polymerization/drug effects , Base Sequence , DNA/genetics , Deoxycytidine/chemistry , Deoxycytidine/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Gemcitabine
8.
ACS Appl Mater Interfaces ; 12(40): 44700-44709, 2020 Oct 07.
Article in English | MEDLINE | ID: mdl-32933250

ABSTRACT

Lead halide perovskite has in recent years gained widespread interest due to its excellent physical and chemical properties, as well as superior optoelectronic performance. However, some restrictions still preclude full industrialization of the material, in particular toxicity issues and instability as a result to sensitivity to humidity. Lead-free all-inorganic double perovskite materials have thus recently become a focus of research. Herein, a new narrow bandgap lead-free double perovskite solar cell with a high-quality Cs2PtI6 film is proposed. It exhibits an optical bandgap of 1.37 eV, absorption within a wide range of wavelengths, and a high absorption coefficient. Following optimization, the device displays a best power conversion efficiency of 0.72% with an open-circuit voltage of 0.73 V, a short-circuit current of 1.2 mA/cm2, and a fill factor of 0.82. Crucially, it also demonstrates excellent stability when exposed to extreme conditions such as high humidity, high temperature, and UV-light irradiation. Stability tests show that the PSCs can retain almost 80% of the original efficiency over 60 days stored in ambient temperature without any encapsulation, boosting prospects for applications of lead-free perovskite solar cells.

9.
Elife ; 92020 05 27.
Article in English | MEDLINE | ID: mdl-32459177

ABSTRACT

Circadian oscillations are generated via transcriptional-translational negative feedback loops. However, individual cells from fibroblast cell lines have heterogeneous rhythms, oscillating independently and with different period lengths. Here we showed that heterogeneity in circadian period is heritable and used a multi-omics approach to investigate underlying mechanisms. By examining large-scale phenotype-associated gene expression profiles in hundreds of mouse clonal cell lines, we identified and validated multiple novel candidate genes involved in circadian period determination in the absence of significant genomic variants. We also discovered differentially co-expressed gene networks that were functionally associated with period length. We further demonstrated that global differential DNA methylation bidirectionally regulated these same gene networks. Interestingly, we found that depletion of DNMT1 and DNMT3A had opposite effects on circadian period, suggesting non-redundant roles in circadian gene regulation. Together, our findings identify novel gene candidates involved in periodicity, and reveal DNA methylation as an important regulator of circadian periodicity.


Subject(s)
Circadian Rhythm/genetics , Epigenesis, Genetic/genetics , Animals , Cells, Cultured , Clone Cells , DNA (Cytosine-5-)-Methyltransferase 1/genetics , DNA (Cytosine-5-)-Methyltransferases/genetics , DNA Methylation/genetics , DNA Methyltransferase 3A , Gene Knockdown Techniques , Gene Regulatory Networks/genetics , Genome/genetics , Humans , Male , Mice , Phenotype , Transcriptome/genetics
10.
ACS Appl Mater Interfaces ; 12(12): 13931-13940, 2020 Mar 25.
Article in English | MEDLINE | ID: mdl-32119775

ABSTRACT

All-inorganic lead halide perovskite solar cells (PSCs) have drawn widespread interest because of its excellent thermal stability compared to its organic-inorganic hybrid counterpart. Poor phase stability caused by moisture, however, has thus far limited their commercial application. Herein, by modifying the interface between the hole-transport layer (HTL) and the perovskite light absorption layer, and by optimizing the HTL for better energy alignment, we controlled the growth of perovskite, reduced carrier recombination, facilitated carrier injection and transport, and improved the PSC's power conversion efficiency (PCE) and moisture stability. When testing using a positive bias scan, we obtained a significant improvement in PCE, 9.49%, which is the champion efficiency of CsPbIBr2-based inverted PSC at present. The stability measurement shows that the passivated CsPbIBr2-based inverted PSCs can retain 86% of its initial efficiency after 1000 h preserved in ambient air with 65% relative humidity. This study paves a new way for enhancing the moisture stability and power conversion efficiency of CsPbIBr2-based PSCs.

11.
Proc Natl Acad Sci U S A ; 116(44): 22229-22236, 2019 10 29.
Article in English | MEDLINE | ID: mdl-31611405

ABSTRACT

Fumarylacetoacetate hydrolase (FAH) is the last enzyme in tyrosine catabolism, and mutations in the FAH gene are associated with hereditary tyrosinemia type I (HT1 or TYRSN1) in humans. In a behavioral screen of N-ethyl-N-nitrosourea mutagenized mice we identified a mutant line which we named "swingshift" (swst, MGI:3611216) with a nonsynonymous point mutation (N68S) in Fah that caused age-dependent disruption of sleep-wake patterns. Mice homozygous for the mutation had an earlier onset of activity (several hours before lights off) and a reduction in total activity and body weight when compared with wild-type or heterozygous mice. Despite abnormal behavioral entrainment to light-dark cycles, there were no differences in the period or phase of the central clock in mutant mice, indicating a defect downstream of the suprachiasmatic nucleus. Interestingly, these behavioral phenotypes became milder as the mice grew older and were completely rescued by the administration of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione], an inhibitor of 4-hydroxyphenylpyruvate dioxygenase, which is upstream of FAH. Mechanistically, the swst mutation had no effect on the enzymatic activity of FAH, but rather promoted the degradation of the mutant protein. This led to reduced FAH protein levels and enzymatic activity in the liver and kidney (but not the brain or fibroblasts) of homozygous mice. In addition, plasma tyrosine-but not methionine, phenylalanine, or succinylacetone-increased in homozygous mice, suggesting that swst mutants provide a model of mild, chronic HT1.


Subject(s)
Circadian Rhythm , Hydrolases/genetics , Mutation , Sleep , Tyrosinemias/genetics , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Animals , Cells, Cultured , Cyclohexanones/therapeutic use , Enzyme Inhibitors/therapeutic use , Enzyme Stability , HEK293 Cells , Homozygote , Humans , Hydrolases/deficiency , Hydrolases/metabolism , Kidney/metabolism , Liver/metabolism , Male , Mice , Mice, Inbred C57BL , Nitrobenzoates/therapeutic use , Organ Specificity , Suprachiasmatic Nucleus/metabolism , Tyrosinemias/drug therapy , Tyrosinemias/physiopathology
12.
ACS Appl Mater Interfaces ; 11(22): 19994-20003, 2019 Jun 05.
Article in English | MEDLINE | ID: mdl-31083899

ABSTRACT

All-inorganic perovskites are attracting increasing attention due to their superior thermal stability than that of the traditional CH3NH3PbI3, while their inferior phase stability in ambient conditions is still an unsolved issue. Here, for the first time, we report the incorporation of niobium (Nb5+) ions into the CsPbI2Br perovskite. Results indicate that Nb5+ can effectively stabilize the photoactive α-CsPbI2Br phase by the possible substitution of Pb2+. With 0.5% Nb doping, the carbon electrode-based all-inorganic perovskite solar cells achieved a high photoconversion efficiency value of 10.42%, 15% higher than that of the control device. The Nb5+ incorporation reduces the charge recombination in the perovskite, leading to a champion Voc value of 1.27 V and a negligible hysteresis effect. This work explicates the high compatibility of all-inorganic perovskite materials and unlocks the opportunities for the use of high-valence ions for perovskite property modification.

13.
RSC Adv ; 9(59): 34152-34157, 2019 Oct 23.
Article in English | MEDLINE | ID: mdl-35529991

ABSTRACT

The high cost of hole transporting materials (HTMs) and noble metal electrodes limits the application of perovskite solar cells (PSCs). Carbon materials have been commonly utilized for HTMs and noble-metal-free PSCs. In this paper, a more conductive 2D MXene material (Ti3C2), showing a similar energy level to carbon materials, has been used as a back electrode in HTMs and noble-metal-free PSCs for the first time. Seamless interfacial contact between the perovskite layer and Ti3C2 material was obtained using a simple hot-pressing method. After the adjustment of key parameters, the PSCs based on the Ti3C2 electrode show more stability and higher power conversion efficiencies (PCE) (13.83%, 27% higher than that (10.87%) of the PSCs based on carbon electrodes) due to the higher conductivity and seamless interfacial contact of the MXene electrode. Our work proposes a promising future application for MXene and also a good electrode candidate for HTM and the noble-metal-free PSCs.

14.
Sci Rep ; 8(1): 6097, 2018 04 17.
Article in English | MEDLINE | ID: mdl-29666396

ABSTRACT

Plants utilize energy from sunlight to perform photosynthesis in chloroplast, an organelle that could be damaged by solar UV radiation. The ultraviolet-B (UV-B) photoreceptor UVR8 is required for UV-B perception and signal transduction. However, little is known about how UVR8 influence chloroplast development under UV-B radiation. Here, we characterized tomato UVR8 gene (SlUVR8) and our results indicated that SlUVR8 facilitate plant acclimation to UV-B stress by orchestrating expression of the UVB-responsive genes (HY5 and CHS) and accumulating UV-absorptive compounds. In addition, we also discovered that SlUVR8 promotes fruit chloroplast development through enhancing accumulation of transcription factor GOLDEN2-LIKE2 (SlGLK2) which determines chloroplast and chlorophyll levels. Furthermore, UV-B radiation could increase expression of SlGLK2 and its target genes in fruits and leaves. SlUVR8 is required for UVB-induced SlGLK2 expression. Together, our work not only identified the conserved functions of SlUVR8 gene in response to UV-B stress, but also uncovered a novel role that SlUVR8 could boost chloroplast development by accumulating SlGLK2 proteins.


Subject(s)
Gene Expression Regulation, Plant/radiation effects , Photoreceptors, Plant/genetics , Plant Proteins/genetics , Solanum lycopersicum/genetics , Transcription Factors/genetics , Ultraviolet Rays/adverse effects , Acclimatization , Chloroplasts/genetics , Chloroplasts/metabolism , Chloroplasts/radiation effects , Fruit/genetics , Fruit/growth & development , Fruit/metabolism , Fruit/radiation effects , Solanum lycopersicum/growth & development , Solanum lycopersicum/metabolism , Solanum lycopersicum/radiation effects , Photoreceptors, Plant/metabolism , Photosynthesis , Plant Proteins/metabolism , Transcription Factors/metabolism
15.
J Biol Chem ; 291(46): 23906-23914, 2016 Nov 11.
Article in English | MEDLINE | ID: mdl-27645993

ABSTRACT

It is known that there are mechanistic links between circadian clocks and metabolic cycles. Reduced nicotinamide adenine dinucleotide (NADH) is a key metabolic cofactor in all living cells; however, it is not known whether levels of NADH oscillate or not. Here we employed REX, a bacterial NADH-binding protein, fused to the VP16 activator to convert intracellular endogenous redox balance into transcriptional readouts by a reporter gene in mammalian cells. EMSA results show that the DNA binding activity of both T- and S-REX::VP16 fusions is decreased with a reduced-to-oxidized cofactor ratio increase. Transient and stabilized cell lines bearing the REX::VP16 and the REX binding operator (ROP) exhibit two circadian luminescence cycles. Consistent with these results, NADH oscillations are observed in host cells, indicating REX can act as a NADH sensor to report intracellular dynamic redox homeostasis in mammalian cells in real time. NADH oscillations provide another metabolic signal for coupling the circadian clock and cellular metabolic states.


Subject(s)
Bacterial Proteins , Biosensing Techniques , Circadian Clocks , Herpes Simplex Virus Protein Vmw65 , NAD/metabolism , Recombinant Fusion Proteins/biosynthesis , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , HEK293 Cells , Herpes Simplex Virus Protein Vmw65/biosynthesis , Herpes Simplex Virus Protein Vmw65/genetics , Humans , Oxidation-Reduction , Recombinant Fusion Proteins/genetics
16.
Plant Sci ; 235: 101-10, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25900570

ABSTRACT

CULLIN 4 (CUL4)-DAMAGED DNA binding protein 1 (DDB1)-based ubiquitin E3 ligase modulates diverse cellular processes including repair of damaged genomic DNA. In this study, an uncharacterized gene termed as DDB1-Interacting protein 2 (DDI2) was identified in yeast two-hybrid screening with bait gene DDB1. The co-immunoprecipitation (co-IP) assays further demonstrated that DDI2 is associated with tomato DDB1-CUL4 complex in vivo. It appears that DDI2 encodes an ortholog of proliferating cell nuclear antigen (PCNA). Confocal microscope observation indicated that DDI2-GFP fusion protein was localized in nuclei. The expression of DDI2 gene is constitutive but substantially enhanced by UV-C irradiation. The transgenic tomato plants with overexpression or knockdown of DDI2 gene displayed the increased or decreased tolerance, respectively, to UV-C stress and chemical mutagen cisplatin. The quantitative analysis of UV-induced DNA lesions indicated that the dark repair of DNA damage was accelerated in DDI2 overexpression lines but delayed in knockdown lines. Conclusively, tomato DDI2 gene is required for UV-induced DNA damage repair and plant tolerance to UV stress. In addition, fruits of DDI2 transgenic plants are indistinguishable from that of wild type, regarding fresh weight and nutrient quality. Therefore, overexpression of DDI2 offers a suitable strategy for genetic manipulation of enhancing plant tolerance to UV stress.


Subject(s)
Adaptation, Physiological/genetics , DNA Damage , DNA Repair/genetics , Genes, Plant , Solanum lycopersicum/genetics , Ultraviolet Rays , Cell Nucleus , Cullin Proteins/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Fruit , Solanum lycopersicum/metabolism , Nuclear Proteins/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Proliferating Cell Nuclear Antigen/genetics , Proliferating Cell Nuclear Antigen/metabolism , Protein Binding , Stress, Physiological/genetics , Ubiquitin-Protein Ligases/metabolism
17.
Int J Mol Med ; 35(1): 31-8, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25394920

ABSTRACT

Fibrosarcoma is an aggressive and highly metastatic cancer of the connective tissue, for which effective therapeutic methods are limited. Recently, there has been a renewed interest in small molecular compounds from natural products in the treatment of cancer. In the present study, we investigated the compound, scutellarein, extracted from the perennial herb Scutellaria lateriflora, and it was found to possess anticancer potential. Cell proliferation assay and cell cycle analysis revealed that the proliferation rate of HT1080 human fibrosarcoma cells was significantly suppressed by treatment with scutellarein through the induction of apoptosis. Moreover, an in vivo experiment using Balb/c nude mice revealed that the volume and weight of the tumors were markedly reduced following treatment with scutellarein. We also analyzed the effects of scutellarein on the markers of metastasis, using the HT1080 cells. The results indicated that scutellarein potently inhibited cell migration, invasion and the expression and activity of matrix metalloproteinase (MMP)-2, -9 and -14. Furthermore, MMP activation and cell survival were suppressed due to the scutellarein-mediated downregulation of nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) activation. In conclusion, our data suggest that scutellarein has the ability to attenuate the development of fibrosarcoma and inhibit cancer cell metastasis.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apigenin/pharmacology , Cell Transformation, Neoplastic/drug effects , Fibrosarcoma/pathology , Animals , Antineoplastic Agents, Phytogenic/administration & dosage , Apigenin/administration & dosage , Apoptosis/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Disease Models, Animal , Fibrosarcoma/genetics , Fibrosarcoma/metabolism , Gene Expression , Humans , Male , Matrix Metalloproteinases/genetics , Matrix Metalloproteinases/metabolism , Mice , NF-kappa B/metabolism , Signal Transduction , Tumor Burden/drug effects , Tumor Stem Cell Assay , Xenograft Model Antitumor Assays
18.
Transgenic Res ; 24(2): 309-17, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25271076

ABSTRACT

Gene targeting is a critical tool for construction of disease models. However, the application of traditional homologous recombination-mediated gene knockout technology is limited by the absence of rapid frequency-guaranteed targeting methods. Although conventional small hairpin RNA (shRNA)-mediated gene silencing offers an alternative for gene targeting, its application is frequently compromised by lower expression efficiency via RNA interference compared to gene knockout. Here we provide an efficient gene targeting strategy involving drug-inducible synergistic silencing with multiple shRNA molecules. On induction, the levels of the target proteins decreased to undetectable levels in all the tested stable transgenic mammalian cell lines, including HEK293 and embryonic stem cell-derived progenies carrying shRNA silencing cassettes. In a transgenic mouse model carrying a silencing cassette targeting the rhodopsin gene, short-time inducer treatment was sufficient to ablate the rhodopsin protein in the retina, resulting in similar retinal phenotypic changes as those observed in rhodopsin mutant mice. Therefore, on a broad basis, this inducible shRNA gene targeting strategy offers a true gene knockout alternative comparable to conventional RNA interference approaches.


Subject(s)
Gene Silencing , Gene Targeting/methods , RNA, Small Interfering/genetics , Rhodopsin/genetics , Animals , Cell Line , Embryonic Stem Cells/metabolism , HEK293 Cells , Humans , Mice , Mice, Transgenic , Models, Animal , Rhodopsin/biosynthesis , Transfection
19.
Biochem Biophys Res Commun ; 446(4): 1022-8, 2014 Apr 18.
Article in English | MEDLINE | ID: mdl-24680685

ABSTRACT

The extracellular matrix (ECM) is an essential element of mammalian organisms, and its cross-linking formation plays a vital role in ECM development and postnatal homeostasis. Defects in cross-link formation caused by aging, genetic, or environmental factors are known to cause numerous diseases in mammals. To augment the cross-linking formation of ECM, the present study established a ZsGreen reporter system controlled by the promoter of lysyl oxidase-like 1 gene (LOXL1), which serves as both a scaffold element and a cross-linking enzyme in the ECM. By using this system in a drug screen, we identified emodin as a strong enhancer of LOXL1 expression that promoted cross-linking formation of ECM in all the tested systems, including human fibroblast cells, cultured human skin tissues, and animals that received long-term emodin treatment. Collectively, the results suggest that emodin may serve as an effective drug or supplement for ECM homeostasis.


Subject(s)
Amino Acid Oxidoreductases/metabolism , Emodin/pharmacology , Extracellular Matrix/drug effects , Protein Kinase Inhibitors/pharmacology , Amino Acid Oxidoreductases/genetics , Animals , Cell Line , Desmosine/metabolism , Elastin/metabolism , Extracellular Matrix/metabolism , Homeostasis/drug effects , Humans , Hydroxyproline/metabolism , Promoter Regions, Genetic/drug effects , Up-Regulation
20.
PLoS One ; 8(10): e73673, 2013.
Article in English | MEDLINE | ID: mdl-24098328

ABSTRACT

Human adipose tissue-derived stem cells (ADSCs) are an attractive multipotent stem cell source with therapeutic applicability across diverse fields for the repair and regeneration of acute and chronically damaged tissues. In recent years, there has been increasing interest in ADSC for tissue engineering applications. However, the mechanisms underlying the regulation of ADSC proliferation are not fully understood. Here we show that 47 transcripts are up-regulated while 23 are down-regulated in ADSC compared to terminally differentiated cells based on global mRNA profiling and microRNA profiling. Among the up-regulated genes, the expression of vascular endothelial growth factor (VEGF) is fine-tuned by miR-199a-5p. Further investigation indicates that VEGF accelerates ADSC proliferation whereas the multipotency of ADSC remains stable in terms of adipogenic, chondrogenic and osteogenic potentials after VEGF treatment, suggesting that VEGF may serve as an excellent supplement for accelerating ADSC proliferation during in vitro expansion.


Subject(s)
Adipose Tissue/cytology , Multipotent Stem Cells/cytology , Vascular Endothelial Growth Factor A/metabolism , 3' Untranslated Regions/genetics , Adipogenesis/drug effects , Cell Proliferation/drug effects , Chondrogenesis/drug effects , Gene Expression Profiling , Gene Expression Regulation/drug effects , Genomics , Humans , MicroRNAs/genetics , Multipotent Stem Cells/drug effects , Multipotent Stem Cells/metabolism , Osteogenesis/drug effects , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/pharmacology
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