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https://www.readbyqxmd.com/read/28745324/a-committed-postselection-precursor-to-natural-tcr%C3%AE-%C3%AE-intraepithelial-lymphocytes
#1
Christoph S N Klose, Jonas F Hummel, Lena Faller, Yannick d'Hargues, Karolina Ebert, Yakup Tanriver
The intestine is a major immune organ with several specialized lymphoid structures and immune cells. Among these are thymus-derived natural intraepithelial lymphocytes (IELs) that lack expression of the classical co-receptors CD4 or CD8αβ (double negative (DN)). Natural IELs are both αβ(+) and γδ(+) T cells that play important roles in the maintenance of the epithelial barrier at steady state and during inflammation. The transcription factor T-bet is essential for the peripheral development of natural IELs, but its role during thymic development has remained less clear...
July 26, 2017: Mucosal Immunology
https://www.readbyqxmd.com/read/28681159/selectable-marker-independent-transformation-of-recalcitrant-maize-inbred-b73-and-sorghum-p898012-mediated-by-morphogenic-regulators-baby-boom-and-wuschel2
#2
Muruganantham Mookkan, Kimberly Nelson-Vasilchik, Joel Hague, Zhanyuan J Zhang, Albert P Kausch
Discriminatory co-expression of maize BBM and WUS transcriptional factor genes promoted somatic embryogenesis and efficient Agrobacterium -mediated transformation of recalcitrant maize inbred B73 and sorghum P898012 genotypes without use of a selectable marker gene. The use of morphogenic regulators to overcome barriers in plant transformation is a revolutionary breakthrough for basic plant science and crop applications. Current standard plant transformation systems are bottlenecks for genetic, genomic, and crop improvement studies...
July 5, 2017: Plant Cell Reports
https://www.readbyqxmd.com/read/28617813/identification-of-padi2-as-a-novel-angiogenesis-regulating-gene-by-genome-association-studies-in-mice
#3
Mehrdad Khajavi, Yi Zhou, Amy E Birsner, Lauren Bazinet, Amanda Rosa Di Sant, Alex J Schiffer, Michael S Rogers, Subrahmanian Tarakkad Krishnaji, Bella Hu, Vy Nguyen, Leonard Zon, Robert J D'Amato
Recent findings indicate that growth factor-driven angiogenesis is markedly influenced by genetic variation. This variation in angiogenic responsiveness may alter the susceptibility to a number of angiogenesis-dependent diseases. Here, we utilized the genetic diversity available in common inbred mouse strains to identify the loci and candidate genes responsible for differences in angiogenic response. The corneal micropocket neovascularization assay was performed on 42 different inbred mouse strains using basic fibroblast growth factor (bFGF) pellets...
June 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28597477/effective-expression-of-drebrin-in-hippocampus-improves-cognitive-function-and-alleviates-lesions-of-alzheimer-s-disease-in-app-swe-ps1-%C3%AE-e9-mice
#4
Yan Liu, Yan-Feng Xu, Ling Zhang, Lan Huang, Pin Yu, Hua Zhu, Wei Deng, Chuan Qin
AIMS: Alzheimer's disease (AD), a progressive development dementia, is increasingly impacting patients' living conditions worldwide. Despite medical care and funding support, there are still no highly individualized drugs and practical strategies for clinical prevention and treatment. Developmentally regulated brain protein (abbreviated as Drebrin or Dbn, also known as Dbn1 in mouse) exists in neurons, especially in dendrites, and is an actin-binding protein that modulates synaptic morphology and long-term memory...
July 2017: CNS Neuroscience & Therapeutics
https://www.readbyqxmd.com/read/28584633/construction-of-a-general-albumin-promoter-reporter-system-for-real-time-monitoring-of-the-differentiation-status-of-functional-hepatocytes-from-stem-cells-in-mouse-rat-and-human
#5
Jing Tang, Qiong Wu, Yi Li, Xiujuan Wu, Yujia Wang, Lihua Zhu, Yujun Shi, Hong Bu, Ji Bao, Mingjun Xie
Genetic constructs with promoters fused to reporter genes for simultaneous monitoring of cellular events have been the focus of attention in recent years. Adenoviral vectors, which have distinctive characteristics, have been used to monitor the differentiation of stem cells in vitro. In the present study, a modified adenoviral vector was constructed, containing a mouse, rat, and human general albumin promoter sequence fused to a ZsGreen reporter gene, and evaluated its efficiency in different cell types. Two hepatocyte cell lines (Hepa1-6 and HepG2), rat primary hepatocytes, rat bone marrow mesenchymal stem cells (BM-MSCs) and rat BM-MSCs-derived hepatocyte-like cells were transduced with this vector, and the transfection efficiency and functional capabilities of the promoter were evaluated by fluorescent microscopy...
June 2017: Biomedical Reports
https://www.readbyqxmd.com/read/28583151/identification-and-validation-of-a-schistosoma-japonicum-u6-promoter
#6
Qing Li, Wan Wang, Nan Zhao, Pengcheng Li, Yue Xin, Wei Hu
BACKGROUND: RNA polymerase III promoters have been widely used to express short hairpin-RNA (shRNA), microRNA (miRNA), and small guide RNA (sgRNA) in gene functional analysis in a variety of organisms including Schistosoma mansoni. However, no endogenous RNA polymerase III promoters have been identified in Schistosoma japonicum. The lack of appropriate promoters in S. japonicum has hindered its gene functional analysis. Identification of functional promoters in S. japonicum is therefore in urgent need...
June 5, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28535287/a-flow-cytometry-based-screen-identifies-mbnl1-modulators-that-rescue-splicing-defects-in-myotonic-dystrophy-type-i
#7
Fan Zhang, Nicole E Bodycombe, Keith M Haskell, Yumei L Sun, Eric T Wang, Carl A Morris, Lyn H Jones, Lauren D Wood, Mathew T Pletcher
Myotonic Dystrophy Type 1 (DM1) is a rare genetic disease caused by expansion of CTG trinucleotide repeats ((CTG)exp) in the 3' untranslated region of the DMPK gene. The repeat transcripts sequester the RNA binding protein Muscleblind-like protein 1 (MBNL1) and hamper its normal function in pre-mRNA splicing. Overexpressing exogenous MBNL1 in the DM1 mouse model has been shown to rescue the splicing defects and reverse myotonia. Although a viable therapeutic strategy, pharmacological modulators of MBNL1 expression have not been identified...
May 23, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/28339054/overexpression-of-tim-3-reduces-helicobacter-pylori-associated-inflammation-through-tlr4-nf%C3%AE%C2%BAb-signaling-in%C3%A2-vitro
#8
Fucai Wang, Zhirong Mao, Dongsheng Liu, Jing Yu, Youhua Wang, Wen Ye, Dongjia Lin, Nanjin Zhou, Yong Xie
The present study aimed to investigate the interaction between T-cell immunoglobulin and mucin-domain-containing molecule-3 (Tim-3) and Toll-like receptor 4 (TLR4)/nuclear factor κB (NF‑κB) signaling in Helicobacter pylori-infected RAW264.7 macrophage cells. RAW264.7 cells were co‑cultured with H. pylori SS1 at different bacteria/cell ratios, and subsequently the mRNA expression of Tim‑3, TLR4, and myeloid differentiation factor 88 (MyD88) was measured by reverse transcription-quantitative polymerase chain reaction (RT‑qPCR)...
May 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28251288/rnai-as-a-tool-to-control-the-sex-ratio-of-mouse-offspring-by-interrupting-zfx-zfy-genes-in-the-testis
#9
YongSheng Zhang, JiFeng Xi, Bin Jia, XiangZu Wang, XuHai Wang, ChaoCheng Li, YaQiang Li, XianCun Zeng, RuiWen Ying, Xin Li, Song Jiang, FangYuan Yuan
The objective of this study was to explore a novel method to alter the sex-ratio balance of mouse offspring by silencing the paralogous genes Zfx/Zfy (Zinc finger X/Y-chromosomal transcription factor gene) during spermatogenesis. Four recombined vectors PRZ1, PRZ2, PRZ3, and PRZ4 (RNAi-Ready-pSIREN-RetroQ-ZsGreen) were constructed for interrupting the Zfx gene. Additionally, a recombined vector Psilencer/Zfy-shRNA was constructed for interrupting the Zfy gene. Male mice were randomly divided into 8 groups, with 20 animals per group...
March 1, 2017: Mammalian Genome: Official Journal of the International Mammalian Genome Society
https://www.readbyqxmd.com/read/27906115/established-cell-surface-markers-efficiently-isolate-highly-overlapping-populations-of-skeletal-muscle-satellite-cells-by-fluorescence-activated-cell-sorting
#10
Claire C Maesner, Albert E Almada, Amy J Wagers
BACKGROUND: Fluorescent-activated cell sorting (FACS) has enabled the direct isolation of highly enriched skeletal muscle stem cell, or satellite cell, populations from postnatal tissue. Several distinct surface marker panels containing different positively selecting surface antigens have been used to distinguish muscle satellite cells from other non-myogenic cell types. Because functional and transcriptional heterogeneity is known to exist within the satellite cell population, a direct comparison of results obtained in different laboratories has been complicated by a lack of clarity as to whether commonly utilized surface marker combinations select for distinct or overlapping subsets of the satellite cell pool...
November 8, 2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27826411/established-cell-surface-markers-efficiently-isolate-highly-overlapping-populations-of-skeletal-muscle-satellite-cells-by-fluorescence-activated-cell-sorting
#11
Claire C Maesner, Albert E Almada, Amy J Wagers
BACKGROUND: Fluorescent-activated cell sorting (FACS) has enabled the direct isolation of highly enriched skeletal muscle stem cell, or satellite cell, populations from postnatal tissue. Several distinct surface marker panels containing different positively selecting surface antigens have been used to distinguish muscle satellite cells from other non-myogenic cell types. Because functional and transcriptional heterogeneity is known to exist within the satellite cell population, a direct comparison of results obtained in different laboratories has been complicated by a lack of clarity as to whether commonly utilized surface marker combinations select for distinct or overlapping subsets of the satellite cell pool...
2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27771389/lassa-and-ebola-virus-inhibitors-identified-using-minigenome-and-recombinant-virus-reporter-systems
#12
Stephen R Welch, Lisa Wiggleton Guerrero, Ayan K Chakrabarti, Laura K McMullan, Mike Flint, Gregory R Bluemling, George R Painter, Stuart T Nichol, Christina F Spiropoulou, César G Albariño
Lassa virus (LASV) and Ebola virus (EBOV) infections are important global health issues resulting in significant morbidity and mortality. While several promising drug and vaccine trials for EBOV are ongoing, options for LASV infection are currently limited to ribavirin treatment. A major factor impeding the development of antiviral compounds to treat these infections is the need to manipulate the virus under BSL-4 containment, limiting research to a few institutes worldwide. Here we describe the development of a novel LASV minigenome assay based on the ambisense LASV S segment genome, with authentic terminal untranslated regions flanking a ZsGreen (ZsG) fluorescent reporter protein and a Gaussia princeps luciferase (gLuc) reporter gene...
October 19, 2016: Antiviral Research
https://www.readbyqxmd.com/read/27724921/claudin-4-activity-in-ovarian-tumor-cell-apoptosis-resistance-and-migration
#13
Douglas A Hicks, Carly E Galimanis, Patricia G Webb, Monique A Spillman, Kian Behbakht, Margaret C Neville, Heidi K Baumgartner
BACKGROUND: Claudin-4 is a transmembrane protein expressed at high levels in the majority of epithelial ovarian tumors, irrespective of subtype, and has been associated with tumor cells that are both chemoresistant and highly mobile. The objective of this study was to determine the functional role that claudin-4 plays in apoptosis resistance and migration as well as the therapeutic utility of targeting claudin-4 activity with a small mimic peptide. METHODS: We examined claudin-4 activity in human ovarian tumor cell lines (SKOV3, OVCAR3, PEO4) using in vitro caspase and scratch assays as well as an in vivo mouse model of ovarian cancer...
October 11, 2016: BMC Cancer
https://www.readbyqxmd.com/read/27640883/a-simple-and-cost-saving-phenotypic-drug-susceptibility-testing-of-hiv-1
#14
Yunceng Weng, Ling Zhang, Jianfeng Huang, Jin Zhao, Peifang Luo, Siyuan Bi, Zhengrong Yang, Hai Zhu, Jean-Pierre Allain, Chengyao Li
It is essential to monitor the occurrence of drug-resistant strains and to provide guidance for clinically adapted antiviral treatment of HIV/AIDS. In this study, an individual patient's HIV-1 pol gene encoding the full length of protease and part of the reverse transcriptase was packaged into a modified lentivirus carrying dual-reporters ZsGreen and luciferase. The optimal coefficient of correlation between drug concentration and luciferase activity was optimized. A clear-cut dose-dependent relationship between lentivirus production and luciferase activity was found in the phenotypic testing system...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27492172/flow-cytometry-and-transplantation-based-quantitative-assays-for-satellite-cell-self-renewal-and-differentiation
#15
Robert W Arpke, Michael Kyba
In response to muscle damage, satellite cells proliferate and undertake both differentiation and self-renewal, generating new functional muscle tissue and repopulating this new muscle with stem cells for future injury responses. For many questions relating to the physiological regulation of satellite cells, quantitative readouts of self-renewal and differentiation can be very useful. There is a particular need for a quantitative assay for satellite cell self-renewal that does not rely solely upon sectioning, staining and counting cells in sections...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27394686/translation-of-bdnf-gene-transcripts-with-short-3-utr-in-hippocampal-ca1-neurons-improves-memory-formation-and-enhances-synaptic-plasticity-relevant-signaling-pathways
#16
Man Wang, Dongxue Li, Di Yun, Yinghan Zhuang, Vez Repunte-Canonigo, Pietro Paolo Sanna, Thomas Behnisch
While the brain-derived neurotrophic factor (BDNF) gene and its multiple transcripts have been recognized as a key factor for learning, but the specific involvement of BDNF translated from BDNF transcripts with short-3' untranslated region (short 3' UTR) in learning and memory requires further analysis. In this paper, we present data to show that the transduction of hippocampal CA1 neurons with AAV9-5' UTR-BDNF (short 3' UTR)-IRES-ZsGreen and the subsequent expression of BDNF enhanced the phosphorylation of synaptic plasticity relevant proteins and improved passive avoidance and object location, but not object recognition memory...
July 6, 2016: Neurobiology of Learning and Memory
https://www.readbyqxmd.com/read/27244456/proteasome-activity-is-required-for-the-initiation-of-precancerous-pancreatic-lesions
#17
Takaki Furuyama, Shinji Tanaka, Shu Shimada, Yoshimitsu Akiyama, Satoshi Matsumura, Yusuke Mitsunori, Arihiro Aihara, Daisuke Ban, Takanori Ochiai, Atsushi Kudo, Hiroshi Fukamachi, Shigeki Arii, Yoshiya Kawaguchi, Minoru Tanabe
Proteasome activity is significantly increased in advanced cancers, but its role in cancer initiation is not clear, due to difficulties in monitoring this process in vivo. We established a line of transgenic mice that carried the ZsGreen-degron(ODC) (Gdeg) proteasome reporter to monitor the proteasome activity. In combination with Pdx-1-Cre;LSL-Kras(G12D) model, proteasome activity was investigated in the initiation of precancerous pancreatic lesions (PanINs). Normal pancreatic acini in Gdeg mice had low proteasome activity...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27113041/generation-of-human-embryonic-stem-cell-line-expressing-zsgreen-in-cholinergic-neurons-using-crispr-cas9-system
#18
Jing Zhou, Chencheng Wang, Kunshan Zhang, Yingying Wang, Xi Gong, Yanlu Wang, Siguang Li, Yuping Luo
Lineage specific human embryonic stem cell (hESC) reporter cell line is a versatile tool for biological studies on real time monitoring of differentiation, physiological and biochemical features of special cell types and pathological mechanism of disease. Here we report the generation of ChAT-zsGreen reporter hESC line that express zsGreen under the control of the choline acetyltransferase (ChAT) promoter using CRISPR (Clustered Regularly Interspersed Short Palindromic Repeats)/Cas9 system. We show that the ChAT-zsGreen hESC reporter cell lines retain the features of undifferentiated hESC...
August 2016: Neurochemical Research
https://www.readbyqxmd.com/read/27076646/partial-podocyte-replenishment-in-experimental-fsgs-derives-from-nonpodocyte-sources
#19
Natalya V Kaverina, Diana G Eng, Remington R S Schneider, Jeffrey W Pippin, Stuart J Shankland
The current studies used genetic fate mapping to prove that adult podocytes can be partially replenished following depletion. Inducible NPHS2-rtTA/tetO-Cre/RS-ZsGreen-R reporter mice were generated to permanently label podocytes with the ZsGreen reporter. Experimental focal segmental glomerulosclerosis (FSGS) was induced with a cytotoxic podocyte antibody. On FSGS day 7, immunostaining for the podocyte markers p57, synaptopodin, and podocin were markedly decreased by 44%, and this was accompanied by a decrease in ZsGreen fluorescence...
June 1, 2016: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/26975544/visualizing-estrogen-receptor-%C3%AE-expressing-neurons-using-a-new-er%C3%AE-zsgreen-reporter-mouse-line
#20
Kenji Saito, Yanlin He, Xiaofeng Yan, Yongjie Yang, Chunmei Wang, Pingwen Xu, Antentor Othrell Hinton, Gang Shu, Likai Yu, Qingchun Tong, Yong Xu
BACKGROUND: A variety of biological functions of estrogens, including regulation of energy metabolism, are mediated by neurons expressing estrogen receptor-α (ERα) in the brain. However, complex intracellular processes in these ERα-expressing neurons are difficult to unravel, due to the lack of strategy to visualize ERα-expressing neurons, especially in unfixed brain tissues. RESULTS AND CONCLUSIONS: Here we generated a novel ERα-ZsGreen reporter mouse line in which expression of a green fluorescent reporter protein, ZsGreen, is driven by a 241kb ERα gene promoter...
April 2016: Metabolism: Clinical and Experimental
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