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https://www.readbyqxmd.com/read/28340094/nlrp2-is-a-suppressor-of-nf-%C3%A6-b-signaling-and-hla-c-expression-in-human-trophoblasts
#1
Tamara Tilburgs, Torsten B Meissner, Leonardo M R Ferreira, Arend Mulder, Kiran Musunuru, Junqiang Ye, Jack L Strominger
During human pregnancy, fetal extravillous trophoblasts (EVT) play a key role in the regulation of maternal T cell and NK cell responses. EVT display a unique combination of Human Leukocyte Antigens (HLA); EVT do not express HLA-A and HLA-B, but do express HLA-C, HLA-E and HLA-G. The mechanisms establishing this unique HLA expression pattern have not been fully elucidated. The MHC class I and class II transcriptional activators NLRC5 and CIITA are expressed neither by EVT nor by the EVT model cell line JEG3, which has an MHC expression pattern identical to that of EVT...
March 7, 2017: Biology of Reproduction
https://www.readbyqxmd.com/read/28339300/genetic-manipulation-by-zinc-finger-nucleases-in-rat-induced-pluripotent-stem-cells
#2
Sheng Yang, Shufang Ding, Qianhua Xu, Xiong Li, Qiong Xiong
Induced pluripotent stem cells (iPSCs) have an extensive application in regenerative medicine, pharmaceutical discovery, and basic research. With the recent derivation of rat iPSCs, it is now feasible to apply genetic manipulation in this species. But such tools do not yet exist for many rat strains, especially for disease model rat. The Sprague Dawley (SD) rat is an inbred disease model for hypertension, nephropathy, pulmonary hypertension, depression, and alcohol consumption. In this study, the SD rat iPSCs were generated using lentiviral method...
March 24, 2017: Cellular Reprogramming
https://www.readbyqxmd.com/read/28333914/genome-scale-crispr-cas9-knockout-and-transcriptional-activation-screening
#3
Julia Joung, Silvana Konermann, Jonathan S Gootenberg, Omar O Abudayyeh, Randall J Platt, Mark D Brigham, Neville E Sanjana, Feng Zhang
Forward genetic screens are powerful tools for the unbiased discovery and functional characterization of specific genetic elements associated with a phenotype of interest. Recently, the RNA-guided endonuclease Cas9 from the microbial CRISPR (clustered regularly interspaced short palindromic repeats) immune system has been adapted for genome-scale screening by combining Cas9 with pooled guide RNA libraries. Here we describe a protocol for genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system...
April 2017: Nature Protocols
https://www.readbyqxmd.com/read/28331616/ex-vivo-akt-inhibition-promotes-the-generation-of-potent-cd19car-t-cells-for-adoptive-immunotherapy
#4
Ryan Urak, Miriam Walter, Laura Lim, ChingLam W Wong, Lihua E Budde, Sandra Thomas, Stephen J Forman, Xiuli Wang
BACKGROUND: Insufficient persistence and effector function of chimeric antigen receptor (CAR)-redirected T cells have been challenging issues for adoptive T cell therapy. Generating potent CAR T cells is of increasing importance in the field. Studies have demonstrated the importance of the Akt pathway in the regulation of T cell differentiation and memory formation. We now investigate whether inhibition of Akt signaling during ex vivo expansion of CAR T cells can promote the generation of CAR T cells with enhanced antitumor activity following adoptive therapy in a murine leukemia xenograft model...
2017: Journal for Immunotherapy of Cancer
https://www.readbyqxmd.com/read/28331087/feline-immunodeficiency-virus-evolutionarily-acquires-two-proteins-vif-and-protease-capable-of-antagonizing-feline-apobec3
#5
Rokusuke Yoshikawa, Junko S Takeuchi, Eri Yamada, Yusuke Nakano, Naoko Misawa, Yuichi Kimura, Fengrong Ren, Takayuki Miyazawa, Yoshio Koyanagi, Kei Sato
The interplay between viral and host proteins has been well studied to elucidate virus-host interactions and their relevance to virulence. Mammalian genes encode apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3) proteins, which act as intrinsic restriction factors against lentiviruses. To overcome APOBEC3-mediated anti-viral actions, lentiviruses have evolutionarily acquired an accessory protein, viral infectivity factor (Vif), and Vif degrades host APOBEC3 proteins via a ubiquitin/proteasome-dependent pathway...
March 22, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28330619/efficient-ex%C3%A2-vivo-engineering-and-expansion-of-highly-purified-human-hematopoietic-stem-and-progenitor-cell-populations-for-gene-therapy
#6
Erika Zonari, Giacomo Desantis, Carolina Petrillo, Francesco E Boccalatte, Maria Rosa Lidonnici, Anna Kajaste-Rudnitski, Alessandro Aiuti, Giuliana Ferrari, Luigi Naldini, Bernhard Gentner
Ex vivo gene therapy based on CD34(+) hematopoietic stem cells (HSCs) has shown promising results in clinical trials, but genetic engineering to high levels and in large scale remains challenging. We devised a sorting strategy that captures more than 90% of HSC activity in less than 10% of mobilized peripheral blood (mPB) CD34(+) cells, and modeled a transplantation protocol based on highly purified, genetically engineered HSCs co-infused with uncultured progenitor cells. Prostaglandin E2 stimulation allowed near-complete transduction of HSCs with lentiviral vectors during a culture time of less than 38 hr, mitigating the negative impact of standard culture on progenitor cell function...
March 13, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28326931/overexpression-of-suppressor-of-cytokine-signaling-3-in-dorsal-root-ganglion-attenuates-cancer-induced-pain-in-rats
#7
Jinrong Wei, Meng Li, Dieyu Wang, Hongyan Zhu, Xiangpeng Kong, Shusheng Wang, You-Lang Zhou, Zhong Ju, Guang-Yin Xu, Guo-Qin Jiang
Background Cancer-induced pain (CIP) is one of the most severe types of chronic pain with which clinical treatment remains challenging and the involved mechanisms are largely unknown. Suppressor of cytokine signaling 3 (SOCS3) is an important intracellular protein and provides a classical negative feedback loop, thus involving in a wide variety of processes including inflammation and nociception. However, the role of SOCS3 pathway in CIP is poorly understood. The present study was designed to investigate the role of SOCS3 in dorsal root ganglion (DRG) in the development of CIP...
January 2017: Molecular Pain
https://www.readbyqxmd.com/read/28326698/-construction-of-human-mucosa-oral-epithelial-cell-lines-overexpressing-telomerase-reverse-transcriptase-gene-mediated-by-lentivirus
#8
Zeng Sa, Qin Xiaodong, He Xiangyi, Che Chunxiao, Zhang Xiao, Xie Siyu, Sun Guijun, Wang Lihe
OBJECTIVE: To construct a cell line of oral mucosa epithelial cells that stably express human telomerase reverse transcriptase (hTERT) by lentiviral vectors, approaches for the establishment of stable and efficient immortalized oral mucosa epithelial cell lines were explored. METHODS: Whole RNA was extracted from 293T cells. The hTERT gene was amplified by polymerase chain reaction (PCR) and cloned into the lentiviral vector as pLVX-puro-hTERT. The lentivirus particles were successfully packaged and used to infect primary oral epithelial cells...
October 1, 2016: Hua Xi Kou Qiang Yi Xue za Zhi, Huaxi Kouqiang Yixue Zazhi, West China Journal of Stomatology
https://www.readbyqxmd.com/read/28322785/puromycin-resistant-lentiviral-control-shrna-vector-plko-1-induces-unexpected-cellular-differentiation-of-p19-embryonic-stem-cells
#9
Jyotshna Kanungo
RNA silencing is used as a common method for investigating loss-of-function effects of genes of interest. In mammalian cells, RNA interference (RNAi) or RNA silencing can be achieved by transient siRNA (small or short interfering RNA) transfection or by stable shRNA (short hairpin RNA) systems. Various vectors are used for efficient delivery of shRNA. Lentiviral vectors offer an efficient delivery system for stable and long-term expression of the shRNA in mammalian cells. The widely used lentiviral pLKO.1 plasmid vector is very popular in RNAi studies...
March 16, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28319069/the-branched-chain-amino-acid-transaminase-1-sustains-growth-of-antiestrogen-resistant-and-er%C3%AE-negative-breast-cancer
#10
V Thewes, R Simon, M Hlevnjak, M Schlotter, P Schroeter, K Schmidt, Y Wu, T Anzeneder, W Wang, P Windisch, M Kirchgäßner, N Melling, N Kneisel, R Büttner, U Deuschle, H P Sinn, A Schneeweiss, S Heck, S Kaulfuss, H Hess-Stumpp, J G Okun, G Sauter, A E Lykkesfeldt, M Zapatka, B Radlwimmer, P Lichter, M Tönjes
Antiestrogen-resistant and triple-negative breast tumors pose a serious clinical challenge because of limited treatment options. We assessed global gene expression changes in antiestrogen-sensitive compared with antiestrogen-resistant (two tamoxifen resistant and two fulvestrant resistant) MCF-7 breast cancer cell lines. The branched-chain amino acid transaminase 1 (BCAT1), which catalyzes the first step in the breakdown of branched-chain amino acids, was among the most upregulated transcripts in antiestrogen-resistant cells...
March 20, 2017: Oncogene
https://www.readbyqxmd.com/read/28317860/gene-delivery-to-nile-tilapia-cells-for-transgenesis-and-the-role-of-pi3k-c2%C3%AE-in-angiogenesis
#11
Fernanda Maria Policarpo Tonelli, Samyra Maria Dos Santos Nassif Lacerda, Marcela Santos Procópio, Breno Luiz Sales Lemos, Luiz Renato de França, Rodrigo Ribeiro Resende
Microinjection is commonly performed to achieve fish transgenesis; however, due to difficulties associated with this technique, new strategies are being developed. Here we evaluate the potential of lentiviral particles to genetically modify Nile tilapia cells to achieve transgenesis using three different approaches: spermatogonial stem cell (SSC) genetic modification and transplantation (SC), in vivo transduction of gametes (GT), and fertilised egg transduction (ET). The SC protocol using larvae generates animals with sustained production of modified sperm (80% of animals with 77% maximum sperm fluorescence [MSF]), but is a time-consuming protocol (sexual maturity in Nile tilapia is achieved at 6 months of age)...
March 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28315290/uca1-regulates-the-growth-and-metastasis-of-pancreatic-cancer-by-sponging-mir-135a
#12
Xiaobo Zhang, Feng Gao, Lei Zhou, Huaitao Wang, Gang Shi, Xiaodong Tan
Pancreatic cancer (PC) is a devastating malignant disease with poor prognosis. This study is aimed to investigate the role of urothelial carcinoma associated 1(UCA1) in the progression of PC. Our results revealed that long non-coding RNAs (lncRNAs)-UCA1 was overexpressed in PC tissues compared with adjacent histologically normal tissues. Down-regulated level of UCA1 was also detected in five human pancreatic cancer cell lines (SW1990,BxPC-3, MiaPaCa-2, PANC-1, CAPAN-1) compared with normal pancreatic duct epithelial HPDE cells...
March 7, 2017: Oncology Research
https://www.readbyqxmd.com/read/28314989/inhibition-of-got1-sensitizes-colorectal-cancer-cells-to-5-fluorouracil
#13
Chengyu Hong, Jian Zheng, Xiaoling Li
PURPOSE: Almost all colorectal cancer (CRC) cell lines are known to overexpress aspartate aminotransferase (GOT1), which potentially regulates the intracellular levels of reactive oxygen species (ROS) via the production of NADPH, and supports tumor growth. In our study, the role of GOT1 in the anticancer efficacy of 5-fluorouracil (5-FU) was examined. METHODS: HCT116, SW480, and HT-29 cells were transfected with lentiviral vectors expressing short hairpin RNA (shRNA) against GOT1...
March 17, 2017: Cancer Chemotherapy and Pharmacology
https://www.readbyqxmd.com/read/28303972/lentiviral-vectors-can-be-used-for-full-length-dystrophin-gene-therapy
#14
John R Counsell, Zeinab Asgarian, Jinhong Meng, Veronica Ferrer, Conrad A Vink, Steven J Howe, Simon N Waddington, Adrian J Thrasher, Francesco Muntoni, Jennifer E Morgan, Olivier Danos
Duchenne Muscular Dystrophy (DMD) is caused by a lack of dystrophin expression in patient muscle fibres. Current DMD gene therapy strategies rely on the expression of internally deleted forms of dystrophin, missing important functional domains. Viral gene transfer of full-length dystrophin could restore wild-type functionality, although this approach is restricted by the limited capacity of recombinant viral vectors. Lentiviral vectors can package larger transgenes than adeno-associated viruses, yet lentiviral vectors remain largely unexplored for full-length dystrophin delivery...
March 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28303373/viral-mediated-overexpression-of-the-myelin-transcription-factor-1-myt1-in-the-dentate-gyrus-attenuates-anxiety-and-ethanol-related-behaviors-in-rats
#15
Amine Bahi, Jean-Luc Dreyer
RATIONALE: Myelin Transcription Factor 1 (MyT1), a member of the Zinc Finger gene family, plays a fundamental role in the nervous system. Recent research has suggested that this transcription factor is associated with the pathophysiology of psychiatric disorders including addiction, schizophrenia, and depression. However, the role of MyT1 in anxiety- and ethanol-related behaviors is still unknown. OBJECTIVES: We evaluated the effects of lentiviral-mediated overexpression of MyT1 in the dentate gyrus (DG) on anxiety- and ethanol-related behaviors in rats...
March 16, 2017: Psychopharmacology
https://www.readbyqxmd.com/read/28301970/disclosing-the-parameters-leading-to-high-productivity-of-retroviral-producer-cells-lines-evaluating-random-vs-targeted-integration
#16
Vanessa Sofia Bandeira, Hélio A Tomás, Evren Alici, Manuel J C T Carrondo, Ana Sofia Coroadinha
Gammaretrovirus and lentivirus are the preferred viral vectors to genetically modify T- and NK- cells to be used in immune-cell therapies. The transduction efficiency of hematopoietic and T cells is more efficient using Gibbon ape leukemia virus (GaLV) pseudotyping. In this context gammaretroviral vector producer cells offer competitive higher titers than transient lentiviral vectors productions. The main aim of this work was to identify the key parameters governing GalV pseudotyped gammaretroviral vector productivity in stable producer cells using a retroviral vector expression cassette enabling positive (facilitating cell enrichment) and negative cell selection (allowing cell elimination)...
March 16, 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28301220/periostin-promotes-migration-proliferation-and-differentiation-of-human-periodontal-ligament-mesenchymal-stem-cells
#17
Ziqiang Wu, Wenyong Dai, Pei Wang, Xiaozhen Zhang, Yi Tang, Lin Liu, Qiaona Wang, Ming Li, Chunbo Tang
PURPOSE: Periostin (POSTN) is critical to bone and dental tissue morphogenesis, postnatal development, and maintenance, however, its roles in tissue repair and regeneration mediated by human periodontal ligament mesenchymal stem cells (PDLSCs) remain unclear. The present study was designed to evaluate the effects of POSTN on hPDLSCs in vitro. MATERIALS AND METHODS: hPDLSCs were isolated and characterized by their expression of the cell surface markers CD44, CD90, CD105, CD34, and CD45...
March 16, 2017: Connective Tissue Research
https://www.readbyqxmd.com/read/28298423/rounding-up-sickle-cells-with-gene-therapy
#18
Leah Byrne
A report of a patient treated with ex vivo lentiviral gene transfer to hematopoietic stem cells shows the promise of gene therapy for sickle cell anemia.
March 15, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/28298247/tissue-specific-inhibition-of-urokinase-type-plasminogen-activator-expression-in-the-testes-of-mice-by-inducible-lentiviral-rna-interference-causes-male-infertility
#19
Kai Zhao, Yan Liu, Zhe Xiong, Lian Hu, Cheng-Liang Xiong
Urokinase-type plasminogen activator (uPA) is involved in many physiological processes, including male infertility. To explore the effects of uPA in male reproduction, we constructed an inducible uPA short hairpin RNA (shRNA) system expressed by lentiviral vectors. After proving inhibition of uPA expression in the mouse Sertoli cell line TM4 by 1μgmL-1 doxycycline (Dox), two lentivirus (pLenti4-shRNA and pLenti6/TR) were co-microinjected into mouse testes to produce TetR&shuPA mice model. Though oral gavage by 0...
March 16, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28297751/preparation-of-vsv-g-viral-envelope-glycoprotein-from-pichia-pastoris-enhances-transfection-of-dna-into-animal-cells
#20
Xin Liu, Ying Dong, Jingquan Wang, Long Li, Zhenmin Zhong, Yun-Pan Li, Shao-Jun Chen, Yu-Cai Fu, Wen-Can Xu, Chi-Ju Wei
Vesicular stomatitis virus G glycoprotein (VSV-G) has been widely used for pseudotyping retroviral, lentiviral and artificial viral vectors. The objective of this study was to establish a potential approach for large scale production of VSV-G. To this end, VSV-G was cloned with a N-terminal His-tag into Pichia pastoris expression vector pPIC3.5K. Three clones (Mut(s)) containing the VSV-G expression cassette were identified by PCR. All clones proliferated normally in expansion medium, while the proliferation was reduced significantly under induction conditions...
March 15, 2017: Journal of Microbiology and Biotechnology
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