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1.
J Virol ; 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32878894

RESUMO

Subgroup J avian leukemia virus (ALV-J), belonging to the genus Alpharetrovirus, enters cells through its envelope surface unit (gp85) via specifically recognizing the cellular receptor chicken Na+/H+ exchanger type I (chNHE1), the 28-39 N-terminal residues of which were characterized as the minimal receptor functional domain in our previous studies. In this study, to further clarify the precise organization and properties of the interaction between ALV-J gp85 and chNHE1, we identified the chNHE1-binding domain of ALV-J gp85 using a series of gp85 mutants with segment substitutions and evaluating their effects on chNHE1-binding in protein-cell binding assays. Our results showed that HA substitutions of amino acids (aa) 38-131 (N-terminal of gp85) and aa 159-283 (C-terminal of gp85) significantly inhibited the interaction between gp85 and chNHE1/chNHE1 loop 1. In addition, these HA-substituted chimeric gp85 proteins could not effectively block the entry of ALV-J into chNHE1-expressing cells. Furthermore, analysis of various N-linked glycosylation site and cysteine mutants in gp85 revealed that glycosylation sites (N6 and N11) and cysteines (C3 and C9) were directly involved in receptor-gp85 binding and important for the entry of ALV-J into cells. Taken together, our findings indicated that the bipartite sequence motif, spanning aa 38-131 and aa 159-283, of ALV-J gp85 was essential for binding to chNHE1, with its two N-linked glycosylation sites and two cysteines being important for its receptor-binding function and subsequent viral infection steps.IMPORTANCE Infection of a cell by retroviruses requires the attachment and fusion of the host and viral membranes. The specific adsorption of env surface proteins to cell receptors is a key step in triggering infections and has been the target of antiviral drug screening. ALV-J is an economically important avian pathogen that belongs to the genus Alpharetrovirus and has a wider host range than other ALV subgroups. Our results showed that the amino acids 38-131 of the N-terminal and 159-283 of the C-terminal of ALV-J gp85 controlled the efficiency of gp85 binding to chNHE1 and were critical for viral infection. In addition, the glycosylation sites (N6 and N11) and cysteines (C9 and C9) of gp85 played a crucial role in the receptor binding and viral entry. These findings might help elucidate the mechanism of the entry of ALV-J into host cells and provide antiviral targets for the control of ALV-J.

2.
Mol Med Rep ; 22(4): 2869-2877, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32945377

RESUMO

Long non­coding RNA high expression in hepatocellular carcinoma (lncRNA HEIH) acts as an oncogene in multiple tumors, including hepatocellular carcinoma, colorectal cancer, melanoma and non­small cell lung cancer. However, the role of HEIH in breast cancer remains unknown. The present study focused on the clinical significance and biological function of HEIH in breast cancer. Specifically, the expression levels of HEIH in breast cancer tissues and breast cancer cell lines were investigated. The results indicated high expression levels of HEIH in human breast cancer tissues, and its expression was positively associated with malignancy status and poor disease prognosis. High expression levels of HEIH were detected in the breast cancer cell lines, including MCF­7, SK­BR­3, MDA­MB­231 and MDA­MB­468. These data were consistent with those derived from the in vivo study. Therefore, small interfering RNA was used to knockdown HEIH expression in order to explore whether HEIH exhibits an oncogenic function in breast cancer. Following HEIH knockdown, the proliferative and metastatic activity of MDA­MB­231 cells was decreased, whereas the induction of cell apoptosis was increased. These results suggested the oncogenic role of HEIH in breast cancer and the potential application of HEIH as an index of malignancy and poor prognosis in breast cancer.

3.
BMC Plant Biol ; 20(1): 383, 2020 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-32819279

RESUMO

BACKGROUND: Hydrogen sulphide (H2S) is involved in regulating physiological processes in plants. We investigated how H2S ameliorates iron (Fe) deficiency in soybean (Glycine max L.) seedlings. Multidisciplinary approaches including physiological, biochemical and molecular, and transcriptome methods were used to investigate the H2S role in regulating Fe availability in soybean seedlings. RESULTS: Our results showed that H2S completely prevented leaf interveinal chlorosis and caused an increase in soybean seedling biomass under Fe deficiency conditions. Moreover, H2S decreased the amount of root-bound apoplastic Fe and increased the Fe content in leaves and roots by regulating the ferric-chelate reductase (FCR) activities and Fe homeostasis- and sulphur metabolism-related gene expression levels, thereby promoting photosynthesis in soybean seedlings. In addition, H2S changed the plant hormone concentrations by modulating plant hormone-related gene expression abundances in soybean seedlings grown in Fe-deficient solution. Furthermore, organic acid biosynthesis and related genes expression also played a vital role in modulating the H2S-mediated alleviation of Fe deficiency in soybean seedlings. CONCLUSION: Our results indicated that Fe deficiency was alleviated by H2S through enhancement of Fe acquisition and assimilation, thereby regulating plant hormones and organic acid synthesis in plants.

5.
Mycoses ; 2020 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-32654196

RESUMO

OBJECTIVE: To evaluate the antifungal susceptibility of Sporothrix globosa isolated from Shandong, China, and compare the differences of antifungal activity in vitro between yeast and mycelial phases. METHODS: The in vitro sensitivity of mycelium phase and yeast phase of Sporothrix globosa to anidulafungin, micafungin, caspofungin, 5-flucytosine, posaconazole, voriconazole, itraconazole, fluconazole and amphotericin B was tested by Sensititre™ YeastOne™. The minimum inhibitory concentration (MIC) values of mycelium phase and yeast phase were calculated. SPSS 19.0 software was used to conduct non-parametric rank sum test for MIC values, and P < .05 was considered statistically significant. RESULTS: The mycelium phase and yeast phase were the most sensitive to itraconazole and the least sensitive to fluconazole. The yeast phase of the same strain was more sensitive to itraconazole, voriconazole, posaconazole, micafungin, anidulafungin, caspofungin and 5-fluorouracil, compared with the mycelium (P < .05). However, fluconazole and amphotericin B had no significant difference in mycelium phase and yeast phase. CONCLUSIONS: Itraconazole is the most active antifungal agent in vitro against S globosa. The yeast phase of the same strain is more sensitive than that of the mycelium.

6.
PLoS One ; 15(7): e0235560, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32614907

RESUMO

The present study investigated the effects of four woody forages (Moringa oleifera Lam (MOL), fermented MOL, Folium mori (FM) and fermented FM) on biodiversity and bioactivity of aerobic culturable gut bacteria of tilapia (Oreochromis niloticus) by a traditional culture-dependent method. A total of 133 aerobic culturable isolates were recovered and identified from the gut of tilapia, belonging to 35 species of 12 genera in three bacterial phyla (Firmicutes, Actinobacteria and Proteobacteria). Among them, 6 bacterial isolates of Bacillus baekryungensis, Bacillus marisflavi, Bacillus pumilus, Bacillus methylotrophicus, Proteus mirabilis and Pseudomonas taiwanensis were isolated from all the five experimental groups. The Bray-Curtis analysis showed that the bacterial communities among the five groups displayed obvious differences. In addition, this result of bioactivity showed that approximate 43% of the aerobic culturable gut bacteria of tilapia displayed a distinct anti-bacterial activity against at least one of four fish pathogens Streptococcus agalactiae, Streptococcus iniae, Micrococcus luteus and Vibrio parahemolyticus. Furthermore, Bacillus amyloliquefaciens and Streptomyces rutgersensis displayed strong activity against all four indicator bacteria. These results contribute to our understanding of the intestinal bacterial diversity of tilapia when fed with woody forages and how certain antimicrobial bacteria flourished under such diets. This can aid in the further exploitation of new diets and probiotic sources in aquaculture.


Assuntos
Bactérias/isolamento & purificação , Ciclídeos/microbiologia , Microbioma Gastrointestinal , Aerobiose , Animais , Antibacterianos/farmacologia , Bacillus/efeitos dos fármacos , Bacillus/genética , Bacillus/isolamento & purificação , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/genética , Biodiversidade , Dieta/veterinária , Microbioma Gastrointestinal/efeitos dos fármacos , Intestinos/microbiologia , Testes de Sensibilidade Microbiana , Filogenia , RNA Ribossômico 16S/química , RNA Ribossômico 16S/metabolismo , Streptococcus/efeitos dos fármacos , Streptococcus/genética , Streptococcus/isolamento & purificação
8.
Infect Genet Evol ; 85: 104425, 2020 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-32561296

RESUMO

Since subgroup J avian leukosis virus (ALV-J) was first isolated in the United Kingdom in 1988, it has seriously hindered the development of the poultry industry worldwide. Although cases of ALV-J infection have been reported as early as 2001 in Pakistan, there was no further research on the isolation and molecular characteristics of ALVs. In the present study, we first isolated two ALVs from suspicious clinical samples that were collected from a desi chicken farm in Pakistan. The results of multiplex PCR and indirect immunofluorescent antibody assays confirmed that the two isolates (PK19FA01 and PK19SA01) belonged to ALV-J. The complete genomes of the two isolates were amplified, sequenced, and systematically analyzed. We found that gp85 of PK19FA01 was more similar to that of the prototype strain HPRS103, whereas gp85 of PK19SA01 was more similar to that of American strains. The two isolates contained an intact E element of 147 residues and had a unique 135 bp deletion in the redundant transmembrane of the 3' untranslated region. The U3 region of the two isolates was highly homologous to that of American ALV-J strains. To our knowledge, this is the first report of the isolation, complete genome sequencing, and systematic molecular epidemiological investigation of ALV-J in Pakistan. Our findings could enrich epidemiological data and might contributed to more effective measures to prevent and control avian leukosis in Pakistan.

9.
Med Sci Monit ; 26: e922673, 2020 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32555132

RESUMO

BACKGROUND Cell cycle arrest and autophagy have been demonstrated to be involved in various transforming growth factor (TGF)-ß-mediated phenotype alterations of tubular epithelial cells (TECs) and tubulointerstitial fibrosis. But the relationship between cell cycle arrest and the autophagy induced by TGF-ß has not been explored well. MATERIAL AND METHODS The effects of autophagy inhibition on TGF-ß-induced cell cycle arrest in TECs were explored in vitro. Human kidney-2 (HK-2) cells were stimulated by TGF-ß with or without a combined treatment of autophagy inhibitor chloroquine (CQ) or bafilomycin A1 (Baf). RESULTS Autophagy inhibition by CQ or Baf promotes the suppression of growth in TGF-ß-treated HK-2 cells, as detected by the Cell Counting Kit-8 (CCK-8) method. In addition, CQ or Baf stimulation enhances G1 arrest in TGF-ß treated HK-2 cells, as investigated using propidium iodide (PI) staining and flow cytometry, which was further confirmed by a decrease in the expression of phosphorylated retinoblastoma protein (p-RB) and cyclin-dependent kinase 4 (CDK4). The upregulation of p21 induced by CQ or Baf may mediate an enhanced G1 arrest in TGF-ß treated HK-2 cells. Western blot analysis showed that TGF-ß-induced expression of extracellular matrix fibronectin was notably upregulated in the presence of autophagy inhibitors. CONCLUSIONS Inhibition of autophagy sensitizes the TECs to G1 arrest and proliferation suppression induced by TGF-ß that contributes to the induction of tubulointerstitial fibrosis.

10.
Vet Microbiol ; 244: 108670, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32402334

RESUMO

Inclusion body hepatitis (IBH), hydropericardium syndrome, and gizzard erosion associated with fowl adenovirus (FAdV) infections are reported globally and resulted in significant poultry industry economic losses. In 2018, severe IBH appeared in Pakistan in a 17-week-old layer flock. Subsequently, a FAdV-11 strain (designated as PKFAd18) was isolated from liver samples and identified based on phylogenetic analyses of the serotype-specific L1 region of the capsid hexon gene. There is no complete genome sequence of the Pakistani FAdV-11. This study successfully sequenced the complete genome of PKFAd18. The full genome of PKFAd18 contains 43 840 base pairs (bp) with a G + C content of 53.9 %, which is comparable to other FAdV serotypes. Similar to other FAdV-11 strains, PKFAd18 has only one fiber, while FAdV-1 and FAdV-4 have two fibers. Notably, PKFAd18 showed unique characteristics compared to other FAdV-11 strains. A natural large genomic deletion (1215 bp) appeared in tandem repeat region two, relative to the ON-NP2 strain. Phylogenetic analyses of the PKFAd18 penton gene showed higher homology with FAdV-9, highlighting potential natural recombination between FAdV-11 and FAdV-9. Moreover, the pathogenicity of PKFAd18 studied in specific-pathogen-free chickens showed that PKFAd18 is capable of inducing severe IBH and could be responsible for IBH in Pakistan. Thus, the first complete genome of FAdV-11 in Pakistan was sequenced in this study, which enriches the diversity of knowledge about FAdV-11 and is useful for developing diagnostics and vaccines for IBH induced by FAdV-11 in Pakistan.

11.
Nanoscale ; 12(18): 10380-10389, 2020 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-32373890

RESUMO

The development of intelligent and precise cancer therapy systems that enable accurate diagnosis and specific elimination of cancer cells while protecting normal cells to improve the safety and effectiveness of the treatment is still a challenge. Herein, we report a novel activatable nanodevice for precise cancer therapy. The nanodevice is constructed by adsorbing a DNA duplex probe onto MnO2 nanosheets. After cellular uptake, the DNA duplex probe undergoes telomerase-triggered conformation switching, resulting in a Ce6 "turn-on" signal for the identification of cancer cells. Furthermore, Deoxyribozyme (DNAzyme) is activated to catalyse the cleavage of survivin mRNA, actualizing a precise synergistic therapy in cancer cells involving photodynamic therapy and gene-silencing. The MnO2 nanosheets provide Mn2+ for the DNAzyme and relieve hypoxia to improve the efficiency of the photodynamic therapy. Live cell studies reveal that this nanodevice can diagnose cancer cells and specifically eliminate them without harming normal cells, so making the treatment safer and more effective. The developed DNA-MnO2 nanodevice provides a valuable and general platform for precise cancer therapy.

12.
Vet Microbiol ; 245: 108700, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32456830

RESUMO

Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease (IBD), an important immunosuppressive disease seriously threatening poultry farming worldwide. Since the identification of the classic strain in 1957, variant IBDV, very virulent IBDV, and novel variant IBDV have successively emerged brought severe challenges. Over the years, attenuated, intermediate, and intermediate-plus live vaccines have been developed to control the disease. The coexistence of various strains in flocks increases the probability of homologous recombination, and in this study, a naturally occurring homologous recombination between a novel variant strain and an intermediate vaccine strain of IBDV was first identified. Sequence analyses demonstrated that the IBD16HeN01 strain was a recombinant IBDV incorporating the skeleton of the novel variant IBDV (SHG19-like strain), where the 3' region of segment A (nt 1539-3260) was replaced by an intermediate vaccine strain (W2512-like strain). Pathogenicity experiments indicated that IBD16HeN01 could cause severe bursal lesions and the recombination increased viral pathogenicity to chick embryos compared with the novel variant IBDV. Homologous recombination in IBDV has increased the complexity of disease prevention and control and reminds us that we should use live vaccines more scientifically and cautiously.

13.
Inorg Chem ; 2020 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-32468810

RESUMO

With the reported CO2 activation for the oxidation of benzene to phenol (-ENE → -OL) by the graphitic carbon nitride g-C3N4 (CN) via an artificial photosynthetic route as inspiration, high-valent actinyls (AnmO2)n+ (An = U, Np, Pu; m = VI, V; n = 2, 1) have been introduced for its further modification. Our calculations indicate thermodynamic spontaneity in the feasibility of g-C3N4-(AnmO2)n+ (CN-Anm) formation. The magnificent structural and electronic properties of CN-Anm are utilized for CO2 activation in terms of the rarely studied -ENE → -OL conversion. The calculated free energies show that most steps of the catalytic cycle are favored by CN-Anm complexes. The first step (carbamate formation) is slightly endothermic in all cases, where CN-U is 0.51 eV higher than CN and CN-Pu is -0.01 eV lower. All benzene addition reactions release energy, with that for CN-U being the lowest. The phenolate formation is favored by some actinyl complexes over CN, and CN-U is only 0.23 eV higher. The phenol release (resulting in formamide complexes) and CO desorption are exothermic for all CN-Anm. The overall process suggests the improved catalytic performance of actinyl-modified CN materials, and the slightly depleted uranyl-carbon nitride could be one of the promising catalysts.

14.
Int J Biol Macromol ; 156: 988-996, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32315681

RESUMO

The lignin amine (LA) was exploited to prepare dually N/S-doped carbon (NSC), which was endowed with intriguing porous structure by Fe3O4 template. N and S elements, originating from LA, are doped into the materials. NSC possesses diverse-scale 3D pores. The macropores are made by Fe3O4, which facilitate to produce meso and micro pores on their walls by KOH activation. The sample prepared at 700 °C (NSC-700) is found to have the largest specific surface area (1199 cm2 g-1) and specific capacity (241 F g-1 at current density of 1 A g-1). Its capacity is 260% as high as that of lignin amine carbon (LAC) prepared without adding Fe3O4. Excellent rate performance is unraveled because of possessing 82% specific capacity at 20 A g-1 and 27.2 Wh kg-1 energy density at 10000 W kg-1 power density. Moreover, the specific capacity maintains 95.0% after 3000 cycles, indicating good electrochemical stability. The good electrical performance of NSC-700 is attributed to its interesting electronic properties that are induced by special pore structure. Because of having merits such as high rate performance, long life, large specific capacity and low cost, our NSC is anticipated to be a promising capacitor as electrode material.

15.
Vet Microbiol ; 243: 108620, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32273006

RESUMO

Infectious bursal disease virus (IBDV), the etiological agent of infectious bursal disease (IBD), is a variable RNA virus of Avibirnavirus. Some artificially attenuated vaccine strains of IBDV can adapt to cell culture of chicken embryo fibroblast (CEF) cell or its immortalized cell line DF1 in vitro while wild-type IBDV cannot. In this study, for the first time, a naturally occurring cell-adapted classic strain (genogroup 1) of IBDV named IBD17JL01 was identified in China. Animal experiments showed that IBD17JL01 could severely damage the central immune organ of infected chickens. Sequence analysis of the full-length genome revealed the peculiar molecular characteristics of IBD17JL01 with a few amino acid substitutions that might be involved in cell-tropism, antigenicity, and virulence of IBDV. Identification of this novel strain is beneficial to our understanding of the complexity of the epidemiology of IBDV. And the expansion of viral cell-tropism might increase the potential risk of the reassortment of different IBDVs including the live vaccines.

16.
Comput Biol Med ; 120: 103721, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32250853

RESUMO

BACKGROUND AND OBJECTIVE: Mismatch between invasive mechanical ventilation and the requirements of patients results in patient-ventilator asynchrony (PVA), which is associated with a series of adverse clinical outcomes. Although the efficiency of the available approaches for automatically detecting various types of PVA from the ventilator waveforms is unsatisfactory, the feasibility of powerful deep learning approaches in addressing this problem has not been investigated. METHODS: We propose a 2-layer long short-term memory (LSTM) network to detect two most frequently encountered types of PVA, namely, double triggering (DT) and ineffective inspiratory effort during expiration (IEE), on two datasets. The performance of the networks is evaluated first using cross-validation on the combined dataset, and then using a cross testing scheme, in which the LSTM networks are established on one dataset and tested on the other. RESULTS: Compared with the reported rule-based algorithms and the machine learning models, the proposed 2-layer LSTM network exhibits the best overall performance, with the F1 scores of 0.983 and 0.979 for DT and IEE detection, respectively, on the combined dataset. Furthermore, it outperforms the other approaches in cross testing. CONCLUSIONS: The findings suggest that LSTM is an excellent technique for accurate recognition of PVA in clinics. Such a technique can help detect and correct PVA for a better patient ventilator interaction.

17.
J Exp Clin Cancer Res ; 39(1): 67, 2020 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-32299469

RESUMO

Tumor microenvironment (TME) is the internal environment in which tumor cells survive, consisting of tumor cells, fibroblasts, endothelial cells, and immune cells, as well as non-cellular components, such as exosomes and cytokines. Exosomes are tiny extracellular vesicles (40-160nm) containing active substances, such as proteins, lipids and nucleic acids. Exosomes carry biologically active miRNAs to shuttle between tumor cells and TME, thereby affecting tumor development. Tumor-derived exosomal miRNAs induce matrix reprogramming in TME, creating a microenvironment that is conducive to tumor growth, metastasis, immune escape and chemotherapy resistance. In this review, we updated the role of exosomal miRNAs in the process of TME reshaping.

18.
Comput Math Methods Med ; 2020: 9763826, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32328158

RESUMO

Objective. The deceleration capacity (DC) and acceleration capacity (AC) of heart rate, which are recently proposed variants to the heart rate variability, are calculated from unevenly sampled RR interval signals using phase-rectified signal averaging. Although uneven sampling of these signals compromises heart rate variability analyses, its effect on DC and AC analyses remains to be addressed. Approach. We assess preprocessing (i.e., interpolation and resampling) of RR interval signals on the diagnostic effect of DC and AC from simulation and clinical data. The simulation analysis synthesizes unevenly sampled RR interval signals with known frequency components to evaluate the preprocessing performance for frequency extraction. The clinical analysis compares the conventional DC and AC calculation with the calculation using preprocessed RR interval signals on 24-hour data acquired from normal subjects and chronic heart failure patients. Main Results. The assessment of frequency components in the RR intervals using wavelet analysis becomes more robust with preprocessing. Moreover, preprocessing improves the diagnostic ability based on DC and AC for chronic heart failure patients, with area under the receiver operating characteristic curve increasing from 0.920 to 0.942 for DC and from 0.818 to 0.923 for AC. Significance. Both the simulation and clinical analyses demonstrate that interpolation and resampling of unevenly sampled RR interval signals improve the performance of DC and AC, enabling the discrimination of CHF patients from healthy controls.

19.
Emerg Microbes Infect ; 9(1): 586-596, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32174269

RESUMO

Since 2015, the prevalence of severe hepatitis-hydropericardium syndrome, which is caused by the novel genotype fowl adenovirus serotype 4 (FAdV-4), has increased in China and led to considerable economic losses. The replication cycle of FAdV-4, especially the emerging highly pathogenic novel genotype FAdV-4, remains largely unknown. The adenovirus fibre interacts with the cellular receptor as the initial step in adenovirus (AdV) infection. In our previous studies, the complete genome sequence showed that the fibre patterns of FAdV-4 were distinct from all other AdVs. Here, protein-blockage and antibody-neutralization assays were performed to confirm that the novel FAdV-4 short fibre was critical for binding to susceptible leghorn male hepatocellular (LMH) cells. Subsequently, fibre 1 was used as bait to investigate the receptor on LMH cells via mass spectrometry. The chicken coxsackie and adenovirus receptor (CAR) protein was confirmed as the novel FAdV-4 receptor in competition assays. We further identified the D2 domain of CAR (D2-CAR) as the active domain responsible for binding to the short fibre of the novel FAdV-4. Taken together, these findings demonstrate for the first time that the chicken CAR homolog is a cellular receptor for the novel FAdV-4, which facilitates viral entry by interacting with the viral short fibre through the D2 domain. Collectively, these findings provide an in-depth understanding of the mechanisms of the emerging novel genotype FAdV-4 invasion and pathogenesis.


Assuntos
Adenoviridae/imunologia , Receptores Virais/imunologia , Adenoviridae/genética , Animais , Anticorpos Antivirais/imunologia , Linhagem Celular , Galinhas , Expressão Gênica , Humanos , Receptores Virais/genética , Solubilidade , Proteínas Virais/genética , Proteínas Virais/imunologia
20.
Vet Microbiol ; 242: 108589, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32122593

RESUMO

The CRISPR/CRISPR-associated protein 9 (Cas9) system is a powerful gene-editing tool originally discovered as an integral mediator of bacterial adaptive immunity. Recently, this technology has been explored for its potential utility in providing new and unique treatments for viral infection. Marek's disease virus (MDV) and avian leukosis virus subgroup J (ALV-J), major immunosuppressive viruses, cause significant economic losses to the chicken industry. Here, we evaluated the efficacy of using MDV as a CRISPR/Cas9-delivery system to directly target and disrupt the reverse-transcribed products of the ALV-J RNA genome during its infection cycle in vitro and in vivo. We first screened multiple potential guide RNA (gRNA) target sites in the ALV-J genome and identified several optimized targets capable of effectively disrupting the latently integrated viral genome and providing efficient defense against new infection by ALV-J in cells. The optimal single-gRNAs and Cas9-expression cassettes were inserted into the genome of an MDV vaccine strain. The results indicated that engineered MDV stably expressing ALV-J-targeting CRISPR/Cas9 efficiently resisted ALV-J challenge in host cells. These findings demonstrated the CRISPR/Cas9 system as an effective treatment strategy against ALV-J infection. Furthermore, the results highlighted the potential of MDV as an effective delivery system for CRISPR/Cas9 in chickens.


Assuntos
Leucose Aviária/prevenção & controle , Sistemas CRISPR-Cas , Herpesvirus Galináceo 2/genética , Vacinas contra Doença de Marek/genética , Doenças das Aves Domésticas/prevenção & controle , Animais , Leucose Aviária/virologia , Vírus da Leucose Aviária/genética , Vírus da Leucose Aviária/patogenicidade , Galinhas/virologia , Fibroblastos/virologia , Edição de Genes/métodos , Vetores Genéticos , Genoma Viral , Doenças das Aves Domésticas/virologia , RNA Guia/genética , RNA Viral/genética , Organismos Livres de Patógenos Específicos , Integração Viral
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