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CRISPR/Cas9 cell death

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https://www.readbyqxmd.com/read/29902453/microrna-29a-inhibition-induces-gab1-upregulation-to-protect-ob-6-human-osteoblasts-from-hydrogen-peroxide
#1
Jian-Wei Ruan, Chen Yao, Jin-Yu Bai, Xiao-Zhong Zhou
The present study determines the role of the Gab1 in hydrogen peroxide (H2 O2 )-induced death of human osteoblasts. We show that Gab1 is required for H2 O2 -induced Akt activation to promote osteoblast survival. In OB-6 human osteoblasts, Gab1 silencing (by targeted-shRNA) or complete knockout (by CRISPR-Cas9 KO plasmid) largely attenuated Akt activation by H2 O2 . Gab1-depleted OB-6 cells were more vulnerable to H2 O2 . Conversely, forced over-expression of Gab1 by an adenovirus vector increased Akt activation to protect OB-6 cells from H2 O2 ...
June 11, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29892077/dying-cells-expose-a-nuclear-antigen-cross-reacting-with-anti-pd-1-monoclonal-antibodies
#2
Philipp Metzger, Sabrina V Kirchleitner, Lars M Koenig, Christine Hörth, Sebastian Kobold, Stefan Endres, Max Schnurr, Peter Duewell
Checkpoint molecules such as programmed death 1 (PD-1) dampen excessive T cell activation to preserve immune homeostasis. PD-1-specific monoclonal antibodies have revolutionized cancer therapy, as they reverse tumour-induced T cell exhaustion and restore CTL activity. Based on this success, deciphering underlying mechanisms of PD-1-mediated immune functions has become an important field of immunological research. Initially described for T cells, there is emerging evidence of unconventional PD-1 expression by myeloid as well as tumor cells, yet, with cell-intrinsic functions in various animal tumor models...
June 11, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29891696/-staphylococcus-aureus-targets-the-purine-salvage-pathway-to-kill-phagocytes
#3
Volker Winstel, Dominique Missiakas, Olaf Schneewind
Staphylococcus aureus colonizes large segments of the human population and causes invasive infections due to its ability to escape phagocytic clearance. During infection, staphylococcal nuclease and adenosine synthase A convert neutrophil extracellular traps to deoxyadenosine (dAdo), which kills phagocytes. The mechanism whereby staphylococcal dAdo intoxicates phagocytes is not known. Here we used CRISPR-Cas9 mutagenesis to show that phagocyte intoxication involves uptake of dAdo via the human equilibrative nucleoside transporter 1, dAdo conversion to dAMP by deoxycytidine kinase and adenosine kinase, and signaling via subsequent dATP formation to activate caspase-3-induced cell death...
June 11, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29891675/specific-binding-of-pcbp1-to-heavily-oxidized-rna-to-induce-cell-death
#4
Takashi Ishii, Hiroshi Hayakawa, Tatsuhiro Igawa, Takeshi Sekiguchi, Mutsuo Sekiguchi
In aerobically growing cells, the guanine base of RNA is oxidized to 8-oxo-7,8-dihydroguanine (8-oxoG), which induces alteration in their gene expression. We previously demonstrated that the human AUF1 protein binds to 8-oxoG in RNA to induce the selective degradation of oxidized messenger RNA. We herein report that the poly(C)-binding protein PCBP1 binds to more severely oxidized RNA to activate apoptosis-related reactions. While AUF1 binds to oligoribonucleotides carrying a single 8-oxoG, PCBP1 does not bind to such oligoribonucleotides but instead binds firmly to oligoribonucleotides in which two 8-oxoG residues are located nearby...
June 11, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29884745/a-role-in-apoptosis-regulation-for-rad-51-gene-of-caenorhabditis-elegans
#5
Marcello Germoglio, Adele Adamo
The evolutionary conserved RAD-51 protein is essential for homologous recombination in the germ line as well as homologous repair of DNA double strand breaks in all eukaryotic cells. In the nematode Caenorhabditis elegans the rad-51 gene is transcribed into messenger-RNAs potentially coding three alternative protein isoforms. Null rad-51 alleles display embryonic lethality, severe defects in chromosome structure and high level of germ line apoptosis. To dissect its functions, we genetically modified the C. elegans rad-51 gene by CRISPR/Cas9 genome-editing technology, obtaining a separation-of-function (sfi-) mutant allele that only disrupts the long transcript isoform...
June 8, 2018: Genetics
https://www.readbyqxmd.com/read/29863336/multiplexed-crispr-cpf1-mediated-genome-editing-in-clostridium-difficile-toward-the-understanding-of-pathogenesis-of-c-difficile-infection
#6
Wei Hong, Jie Zhang, Guzhen Cui, Luxin Wang, Yi Wang
Clostridium difficile is often the primary cause of nosocomial diarrhea, leading to thousands of deaths annually worldwide. The availability of an efficient genome editing tool for C. difficile is essential to understanding its pathogenic mechanism and physiological behavior. Although CRISPR-Cas9 has been extensively employed for genome engineering in various organisms, large gene deletion and multiplex genome editing is still challenging in microorganisms with underdeveloped genetic engineering tools. Here, we describe a streamlined CRISPR-Cpf1-based toolkit to achieve precise deletions of fur, tetM, and ermB1/2 in C...
June 4, 2018: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29849821/crispr-cas9-genome-editing-of-epidermal-growth-factor-receptor-sufficiently-abolished-oncogenicity-in-anaplastic-thyroid-cancer
#7
Li-Chi Huang, Ka-Wai Tam, Wei-Ni Liu, Chun-Yu Lin, Kai-Wen Hsu, Wen-Shyang Hsieh, Wei-Ming Chi, Ai-Wei Lee, Jinn-Moon Yang, Ching-Ling Lin, Chia-Hwa Lee
Anaplastic carcinoma of the thyroid (ATC), also called undifferentiated thyroid cancer, is the least common but most aggressive and deadly thyroid gland malignancy of all thyroid cancers. The aim of this study is to explore essential biomarker and use CRISPR/Cas9 with lentivirus delivery to establish a gene-target therapeutic platform in ATC cells. At the beginning, the gene expression datasets from 1036 cancers from CCLE and 8215 tumors from TCGA were collected and analyzed, showing EGFR is predominantly overexpressed in thyroid cancers than other type of cancers ( P = 0...
2018: Disease Markers
https://www.readbyqxmd.com/read/29778644/a-trichostatin-a-tsa-sp1-mediated-mechanism-for-the-regulation-of-sall2-tumor-suppressor-in-jurkat-t-cells
#8
Matías I Hepp, David Escobar, Carlos Farkas, Viviana Hermosilla, Claudia Álvarez, Roberto Amigo, José L Gutiérrez, Ariel F Castro, Roxana Pincheira
SALL2 is a transcription factor involved in development and disease. Deregulation of SALL2 has been associated with cancer, suggesting that it plays a role in the disease. However, how SALL2 is regulated and why is deregulated in cancer remain poorly understood. We previously showed that the p53 tumor suppressor represses SALL2 under acute genotoxic stress. Here, we investigated the effect of Histone Deacetylase Inhibitor (HDACi) Trichostatin A (TSA), and involvement of Sp1 on expression and function of SALL2 in Jurkat T cells...
May 17, 2018: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29778503/autoinflammatory-mutation-in-nlrc4-reveals-an-lrr-lrr-oligomerization-interface
#9
Fiona Moghaddas, Ping Zeng, Yuxia Zhang, Heike Schützle, Sebastian Brenner, Sigrun R Hofmann, Reinhard Berner, Yuanbo Zhao, Bingtai Lu, Xiaoyun Chen, Li Zhang, Suyun Cheng, Stefan Winkler, Kai Lehmberg, Scott W Canna, Peter E Czabotar, Ian P Wicks, Dominic De Nardo, Christian M Hedrich, Huasong Zeng, Seth L Masters
BACKGROUND: Monogenic autoinflammatory disorders are characterised by dysregulation of the innate immune system, for example by gain-of-function mutations in inflammasome forming proteins such as NLRC4. OBJECTIVE: Here we investigate the mechanism by which a novel mutation in the leucine rich repeat (LRR) domain of NLRC4 (c.G1965C, p.W655C) contributes to autoinflammatory disease. METHODS: We studied two unrelated patients with early onset macrophage activation syndrome (MAS) harboring the same de novo mutation in NLRC4...
May 17, 2018: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/29772059/the-orphan-g-protein-coupled-receptor-75-signaling-is-activated-by-the-chemokine-ccl5
#10
Simona Dedoni, Lee A Campbell, Brandon K Harvey, Valeria Avdoshina, Italo Mocchetti
The chemokine CCL5 prevents neuronal cell death mediated both by amyloid β, as well as the human immunodeficiency virus (HIV) viral proteins gp120 and Tat. Because CCL5 binds to CCR5, CCR3 and/or CCR1 receptors, it is unclear which of these receptors plays a role in neuroprotection. Indeed, CCL5 also has neuroprotective activity in cells lacking these receptors. CCL5 may bind to a G protein-coupled receptor 75 (GPR75), which encodes for a 540 amino-acid orphan receptor of the Gqα family. In this study, we have used SH-SY5Y human neuroblastoma cells to characterize whether CCL5 could activate a Gq signaling through GPR75...
May 17, 2018: Journal of Neurochemistry
https://www.readbyqxmd.com/read/29769640/hypoxia-induced-microrna-191-contributes-to-hepatic-ischemia-reperfusion-injury-through-the-zonab-cyclin-d1-axis
#11
Wenming Pan, Lin Wang, Xiao-Fei Zhang, Hongji Zhang, Jinxiang Zhang, Guoliang Wang, Peng Xu, Yunwei Zhang, Ping Hu, Xiao-Dong Zhang, Run-Lei Du, Hui Wang
Hepatic ischemia/reperfusion injury (IRI) is a common cause of morbidity and mortality in liver transplantation settings and involves severe cell death and inflammatory responses. MicroRNA-191 has recently been reported to be abnormally expressed in hepatocellular carcinoma and other liver diseases in the regulation of important cellular processes. However, little is known about its function and molecular mechanism in IRI. Here, we demonstrate that miR-191 is significantly upregulated in a cultured cell line during hypoxia/reperfusion (H/R) and in liver tissue during IRI in mice...
May 16, 2018: Cell Death and Differentiation
https://www.readbyqxmd.com/read/29766242/inactivation-of-deubiquitinase-cyld-enhances-therapeutic-antibody-production-in-chinese-hamster-ovary-cells
#12
Yafang Lu, Qin Zhou, Qianqian Han, Pengfei Wu, Lanlan Zhang, Lin Zhu, David T Weaver, Changzhi Xu, Buchang Zhang
Chinese hamster ovary (CHO) cells are promising host engineering cells for industry manufacturing of therapeutic antibodies. However, cell death due to apoptosis remains a huge challenge to augment antibody production, and developing CHO cells with enhanced anti-apoptosis and proliferation ability is fundamental for cell line development and high-yielding bioprocesses. Deubiquitinase cylindromatosis (CYLD) has been proved to be a tumor suppressor by negatively regulating NF-κB and Wnt/β-catenin signaling pathways...
May 15, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29760953/hyperglycemia-potentiates-a-shift-from-apoptosis-to-rip1-dependent-necroptosis
#13
William D McCaig, Payal S Patel, Sergey A Sosunov, Nicole L Shakerley, Tori A Smiraglia, Miranda M Craft, Katharine M Walker, Matthew A Deragon, Vadim S Ten, Timothy J LaRocca
Apoptosis and necroptosis are the primary modes of eukaryotic cell death, with apoptosis being non-inflammatory while necroptosis is highly inflammatory. We previously demonstrated that, once activated, necroptosis is enhanced by hyperglycemia in several cell types. Here, we determine if hyperglycemia affects apoptosis similarly. We show that hyperglycemia does not enhance extrinsic apoptosis but potentiates a shift to RIP1-dependent necroptosis. This is due to increased levels and activity of RIP1, RIP3, and MLKL, as well as decreased levels and activity of executioner caspases under hyperglycemic conditions following stimulation of apoptosis...
2018: Cell Death Discovery
https://www.readbyqxmd.com/read/29733844/pf-06409577-activates-ampk-signaling-to-protect-retinal-pigment-epithelium-cells-from-uv-radiation
#14
Xiao-Feng Li, Xiu-Ming Liu, Da-Rui Huang, Hai-Jing Cao, Jun-Ying Wang
Ultra-violet (UV) radiation (UVR) to human retinas induces oxidative injury to the resident retinal pigment epithelium (RPE) cells. PF-06409577 a novel, potent and direct AMP-activated protein kinase (AMPK) activator. In ARPE-19 cells and primary murine RPE cells, PF-06409577 significantly inhibited UVR-induced viability reduction, cell death and apoptosis. PF-06409577 activated AMPK signaling in RPE cells by increasing AMPKα1-acetyl-CoA carboxylase phosphorylation and AMPK activity. AMPK inhibition, by AMPKα1-shRNA, -CRISPR/Cas9 knockout or -T172A dominant negative mutation, almost abolished PF-06409577-induced RPE cytoprotection against UVR...
May 4, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29709474/keratinocyte-growth-factor-protects-endometrial-cells-from-oxygen-glucose-deprivation-re-oxygenation-via-activating-nrf2-signaling
#15
Xuting Shi, Hia-Yan Liu, Shu-Ping Li, Hong-Bin Xu
Oxygen and glucose deprivation (OGD)-re-oxygenation (OGDR) exposure to endometrial cells mimics ischemia-reperfusion injury. The present study tests the potential effect of keratinocyte growth factor (KGF) on the process. We show that KGF receptor KGFR is expressed in human endometrial T-HESC cells and primary murine endometrial cells. KGF pre-treatment protected endometrial cells from OGDR, inhibiting cell viability reduction and cell death. KGF attenuated OGDR-induced programmed necrosis in endometrial cells...
April 27, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29709426/the-activation-of-stim1-mediates-s-phase-arrest-and-cell-death-in-paraquat-induced-acute-lung-intoxication
#16
Hanting Fan, Huijie Huang, Li Hu, Wenjia Zhu, Yanfang Yu, Jiaqian Lou, Lingli Hu, Feng Chen
Calcium signaling and oxidative stress are tightly linked to cell cycle and cell death in response to a number of stress conditions. Recent study indicated that stromal interaction molecule 1 (STIM1) is the endoplasmic reticulum calcium sensor. However, the regulatory mechanisms and the role of STIM1 in paraquat (PQ)-induced acute lung intoxication remain elusive. The aim of this study was to explore the molecular and cellular mechanisms of PQ induced acute intoxication in the lung, and further determine whether calcium signaling and reactive oxygen species (ROS) participate in the regulatory mechanism...
April 27, 2018: Toxicology Letters
https://www.readbyqxmd.com/read/29686375/requirement-of-gsk-3-for-puma-induction-upon-loss-of-pro-survival-pi3k-signaling
#17
Florian Schubert, Juliane Rapp, Prisca Brauns-Schubert, Lisa Schlicher, Kerstin Stock, Manuela Wissler, Martina Weiß, Céline Charvet, Christoph Borner, Ulrich Maurer
Growth factor withdrawal induces rapid apoptosis via mitochondrial outer membrane permeabilization. We had previously observed that cell death of IL-3-dependent Ba/F3 cells, induced by removal of the growth factor, required the activity of the kinase GSK-3. Employing CRISPR/Cas9-mediated gene knockout, we aimed to identify pro-apoptotic GSK-3 regulated factors in this process. Knockout of either Puma or Bim demonstrated that the induction of Puma, but not Bim, was crucial for apoptosis induced by IL-3 deprivation...
April 23, 2018: Cell Death & Disease
https://www.readbyqxmd.com/read/29672299/crispr-cas9-mediated-silencing-of-cd44-in-human-highly-metastatic-osteosarcoma-cells
#18
Tang Liu, Zuyun Yan, Yong Liu, Edwin Choy, Francis J Hornicek, Henry Mankin, Zhenfeng Duan
BACKGROUND/AIMS: Metastasis is the major cause of death in patients with osteosarcoma. There is an urgent need to identify molecular markers that promote metastasis. Cluster of differentiation 44 is a receptor for hyaluronic acid (HA) and HA-binding has been proven to participate in various biological tumor activities, including tumor progression and metastasis. METHODS: We performed a meta-analysis to investigate the relationship between CD44 expression, survival, and metastasis in patients with osteosarcoma...
2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29667553/car-t-cell-therapy-a-new-era-in-cancer-immunotherapy
#19
Androulla N Miliotou, Lefkothea C Papadopoulou
BACKGROUND: Cancer is one of the leading causes of death worldwide. Over the years, a number of conventional cytotoxic approaches for neoplastic diseases has been developed. However, due to their limited effectiveness in accordance with the heterogeneity of cancer cells, there is a constant search for therapeutic approaches with improved outcome, such as immunotherapy that utilizes and enhances the normal capacity of the patient's immune system. METHODS: Chimeric Antigen Receptor (CAR) T-cell therapy involves genetic modification of patient's autologous T-cells to express a CAR specific for a tumor antigen, following by ex vivo cell expansion and re-infusion back to the patient...
April 17, 2018: Current Pharmaceutical Biotechnology
https://www.readbyqxmd.com/read/29643863/mutation-in-rice-abscisic-acid2-results-in-cell-death-enhanced-disease-resistance-altered-seed-dormancy-and-development
#20
Yongxiang Liao, Que Bai, Peizhou Xu, Tingkai Wu, Daiming Guo, Yongbin Peng, Hongyu Zhang, Xiaoshu Deng, Xiaoqiong Chen, Ming Luo, Asif Ali, Wenming Wang, Xianjun Wu
Lesion mimic mutants display spontaneous cell death, and thus are valuable for understanding the molecular mechanism of cell death and disease resistance. Although a lot of such mutants have been characterized in rice, the relationship between lesion formation and abscisic acid (ABA) synthesis pathway is not reported. In the present study, we identified a rice mutant, lesion mimic mutant 9150 ( lmm9150 ), exhibiting spontaneous cell death, pre-harvest sprouting, enhanced growth, and resistance to rice bacterial and blast diseases...
2018: Frontiers in Plant Science
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