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https://www.readbyqxmd.com/read/29786554/a-human-ipsc-line-capable-of-differentiating-into-functional-macrophages-expressing-zsgreen-a-tool-for-the-study-and-in-vivo-tracking-of-therapeutic-cells
#1
Martha Lopez-Yrigoyen, Antonella Fidanza, Luca Cassetta, Richard A Axton, A Helen Taylor, Jose Meseguer-Ripolles, Anestis Tsakiridis, Valerie Wilson, David C Hay, Jeffrey W Pollard, Lesley M Forrester
We describe the production of a human induced pluripotent stem cell (iPSC) line, SFCi55-ZsGr, that has been engineered to express the fluorescent reporter gene, ZsGreen, in a constitutive manner. The CAG-driven ZsGreen expression cassette was inserted into the AAVS1 locus and a high level of expression was observed in undifferentiated iPSCs and in cell lineages derived from all three germ layers including haematopoietic cells, hepatocytes and neurons. We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsGreen iPSC line and show that they are indistinguishable from those generated from their parental SFCi55 iPSC line in terms of gene expression, cell surface marker expression and phagocytic activity...
July 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29764836/subcellular-localization-of-antigen-dictates-delivery-of-cd4-t-cell-help-for-the-ctl-response-upon-therapeutic-dna-vaccination-into-the-skin
#2
Nikolina Bąbała, Astrid Bovens, Evert de Vries, Victoria Iglesias-Guimarais, Tomasz Ahrends, Matthew F Krummel, Jannie Borst, Adriaan D Bins
In a mouse model of therapeutic DNA vaccination, we studied how the subcellular localization of vaccine protein impacts antigen delivery to professional antigen presenting cells (pAPCs) and efficiency of CTL priming. Cytosolic, membrane-bound, nuclear, and secretory versions of ZsGreen fluorescent protein, conjugated to MHC class I and -II ovalbumin (OVA) epitopes were expressed in keratinocytes by DNA vaccination into the skin. ZsGreen-OVA versions reached B cells in the skin-draining lymph node (dLN) that proved irrelevant for CTL priming...
May 15, 2018: Cancer Immunology Research
https://www.readbyqxmd.com/read/29676999/sex-differences-in-transcriptomic-profiles-in-aged-kidney-cells-of-renin-lineage
#3
Yuliang Wang, Diana G Eng, Jeffrey W Pippin, Sina A Gharib, Aaron McClelland, Kenneth W Gross, Stuart J Shankland
Renin expressing cells in the kidney's juxta-glomeruluar compartment likely also serve as progenitors for adult glomerular cells in disease. Although these cells of renin lineage (CoRL) decrease in number with advancing kidney age, accompanied by less responsiveness to typical stimuli such as ACE-inhibition, mechanisms and the impact of sex as a biological variable with age are not known. Accordingly, labeled CoRL were sorted from individual young (2m) and aged (27m) male and female Ren1cCre|ZsGreen reporter mice, and their transcriptomic profiles analyzed by RNA seq...
April 18, 2018: Aging
https://www.readbyqxmd.com/read/29567386/the-grueneberg-olfactory-organ-neuroepithelium-recovers-after-injury
#4
Fatemeh Chehrehasa, Angela Jacques, James A St John, Jenny A K Ekberg
The Grueneberg organ (also termed Grueneberg ganglion) is an olfactory subsystem at the rostral nasal septum of rodents, and has been suggested to exist also in humans. Grueneberg organ neurons respond to coldness and alarm pheromones, but the anatomical arrangement and regenerative capacity are not fully characterised. We examined the relationship between the glia and the neurons using crosses of two transgenic mouse lines, S100ß-DsRed and OMP-ZsGreen, to visualise olfactory ensheathing cells (OECs) and Grueneberg olfactory neurons, respectively...
March 19, 2018: Brain Research
https://www.readbyqxmd.com/read/29414419/generation-of-a-human-embryonic-stem-cell-line-expressing-tetrameric-zoanthus-sp-green-fluorescent-protein-nerce002-a-1
#5
Xingxiang Duan, Menghan Xie, Yingying Peng, Liang Hu, Juan Yu, Sicong Zeng, Yang Wang, Guangxiu Lu, Ge Lin, Yi Sun
The human embryonic stem cell (hESC) line NERCe002-A-1 was generated through lentiviral transduction of the original NERCe002-A-1 hESC line with Zoanthus sp. green fluorescent protein (ZsGreen). Cells that expressed ZsGreen showed a >8.6-fold increase in fluorescence intensity compared with that of cells that expressed enhanced green fluorescent protein. The fluorescent hESC line can aid in identification of biological characteristics in vitro and in vivo by tracking cell growth, migration, and differentiation...
April 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29344663/adeno-associated-virus-type-2-mediated-gene-transfer-of-a-short-hairpin-rna-targeting-human-igfbp-2-suppresses-the-proliferation-and-invasion-of-mda-mb-468-cells
#6
Chao Gao, Ru-Song Zhang, Nan Zheng, Chen Wang
Adeno-associated virus 2 (AAV2) is prepotent in the biological treatment of breast tumor because of its low pathogenicity and immunogenicity. Our previous study demonstrated that insulin‑like growth factor‑binding protein 2 (IGFBP‑2) was highly expressed in patients with breast metastasis. In the present study, the effects of recombinant AAV2 on the growth and metastasis of breast cancer cells were determined in vitro, and in vivo. rAAV2-ZsGreen-shRNA-scramble and rAAV2‑ZsGreen‑shRNA‑hIGFBP‑2 were used to transfect MDA‑MB‑468, and MCF‑10A cells respectively, and observed that these virus could not penetrate the normal human breast epithelia MCF‑10A cell line...
January 16, 2018: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29276753/indian-hedgehog-suppresses-a-stromal-cell-driven-intestinal-immune-response
#7
B Florien Westendorp, Nikè V J A Büller, Olga N Karpus, Willemijn A van Dop, Jan Koster, Rogier Versteeg, Pim J Koelink, Clinton Y Snel, Sander Meisner, Joris J T H Roelofs, Anja Uhmann, Emiel Ver Loren van Themaat, Jarom Heijmans, Heidi Hahn, Vanesa Muncan, Manon E Wildenberg, Gijs R van den Brink
Background & Aims: Upon intestinal epithelial damage a complex wound healing response is initiated to restore epithelial integrity and defend against pathogenic invasion. Epithelium-derived Indian Hedgehog (Ihh) functions as a critical sensor in this process. Signaling occurs in a paracrine manner because the receptor for Ihh is expressed only in the mesenchyme, but the exact Hedgehog target cell has remained elusive. The aim of this study was to elucidate further the nature of this target cell in the context of intestinal inflammation...
2018: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/29232382/charting-the-transcriptional-landscape-of-cells-of-renin-lineage-following-podocyte-depletion
#8
Aaron D McClelland, Julia Lichtnekert, Diana G Eng, Jeffrey W Pippin, Kenneth W Gross, Sina A Gharib, Stuart J Shankland
Renin producing cells of the juxtaglomerulus, herein called cells of renin lineage (CoRL), have garnered recent interest for their propensity to act as a progenitor source for various kidney cell types including podocytes. Despite recent advances, the process of transdifferentiation of CoRL to podocytes is poorly understood. In this study, we employed a transgenic reporter mouse line which permanently labels CoRL with ZsGreen fluorescent protein, allowing for isolation by fluorescence-activated cell sorting...
2017: PloS One
https://www.readbyqxmd.com/read/29152658/hepatic-differentiation-of-mouse-bone-marrow%C3%A2-derived-mesenchymal-stem-cells-using-a-novel-3d-culture-system
#9
Qiong Wu, Jing Tang, Yi Li, Li Li, Yujia Wang, Ji Bao, Hong Bu
The development of novel culture systems that mimic the in vivo microenvironment may be beneficial for inducing the differentiation of stem cells and promoting liver function. In the present study, spheroid cultures and decellularized liver scaffolds (DLSs) were utilized to obtain differentiated hepatocyte‑like cells. Mouse bone marrow (BM)‑derived mesenchymal stem cells (MSCs) self‑aggregated into spheroids under low‑attachment conditions and implanted into the DLSs via a negative pressure suction device...
December 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29131531/modifying-expression-modes-of-human-neurotensin-receptor-type-1-alters-sensing-capabilities-for-agonists-in-yeast-signaling-biosensor
#10
Hiroki Hashi, Yasuyuki Nakamura, Jun Ishii, Akihiko Kondo
Neurotensin receptor type 1 (NTSR1), a member of the G-protein-coupled receptor (GPCR) family, is naturally activated by binding of a neurotensin peptide, leading to a variety of physiological effects. The budding yeast Saccharomyces cerevisiae is a proven host organism for assaying the agonistic activation of human GPCRs. Previous studies showed that yeast cells can functionally express human NTSR1 receptor, permitting the detection of neurotensin-promoted signaling using a ZsGreen fluorescent reporter gene...
November 13, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/29097083/knockdown-of-melatonin-receptor-1-and-induction-of-follicle-stimulating-hormone-on-the-regulation-of-mouse-granulosa-cell-function
#11
Hira Sajjad Talpur, Tesfaye Worku, Zia Ur Rehman, Rahim Dad, Dinesh Bhattarai, Iqra Bano, Farmanullah, Aixin Liang, Changjiu He, Liguo Yang
Melatonin receptor 1 (MT1) performs a critical role in the regulation of the animal reproductive system, particularly in follicular growth, and has a considerable effect on reproductive performance. However, the role that MT1 plays in regulating hormones associated with reproduction remains unclear. This study was designed to examine the physiological role of constitutive MT1 silencing and follicle stimulating hormone (FSH) treatment in reproduction, making use of mouse granulosa cells (mGCs) as a model. To understand the constitutive role of MT1 in ovarian physiology, the RNAi-Ready pSIREN-RETROQ-ZsGreen Vector mediated recombinant pshRNA was used to silence MT1 gene expression...
December 2017: Reproductive Biology
https://www.readbyqxmd.com/read/29016733/endothelial-to-haematopoietic-transition-contributes-to-pulmonary-arterial-hypertension
#12
Olin D Liang, Eui-Young So, Pamela C Egan, Laura R Goldberg, Jason M Aliotta, Keith Q Wu, Patrycja M Dubielecka, Corey E Ventetuolo, Anthony M Reginato, Peter J Quesenberry, James R Klinger
Aims: The pathogenic mechanisms of pulmonary arterial hypertension (PAH) remain unclear, but involve dysfunctional endothelial cells (ECs), dysregulated immunity and inflammation in the lung. We hypothesize that a developmental process called endothelial to haematopoietic transition (EHT) contributes to the pathogenesis of pulmonary hypertension (PH). We sought to determine the role of EHT in mouse models of PH, to characterize specific cell types involved in this process, and to identify potential therapeutic targets to prevent disease progression...
November 1, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/28879651/preliminary-studies-on-the-differential-expression-of-adrenocorticotropic-hormone-receptor-in-adrenal-adenomas
#13
Dongliang Hu, Tongzu Liu, Xinghuan Wang
INTRODUCTION: The ACTH receptor (ACTHR) is primarily expressed in the adrenal cortex. Previous studies focused on the regulatory function of ACTHR in glucocorticoid secretion, but research on adrenal tumours is rare. The aim of this study was to evaluate ACTHR expression in common adrenal adenomas and investigate its influence on adrenal tumorigenesis using adrenocortical H295R cells. MATERIALS AND M: ETHODS: Real-time polymerase chain reaction and western blot were used to detect the expression of ACTHR in 18 aldosterone-producing adenomas, 16 cortisol-producing adenomas, nine non-functional adenomas, and 12 normal adrenal samples...
September 7, 2017: Endokrynologia Polska
https://www.readbyqxmd.com/read/28745324/a-committed-postselection-precursor-to-natural-tcr%C3%AE-%C3%AE-intraepithelial-lymphocytes
#14
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
#15
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...
September 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
#16
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
#17
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
#18
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
#19
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
#20
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 the 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...
August 15, 2017: Human Molecular Genetics
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