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lentivirus genetic engineering

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https://www.readbyqxmd.com/read/29524405/dual-expression-of-cxcr4-and-il-35-enhances-the-therapeutic-effects-of-bmscs-on-tnbs-induced-colitis-rats-through-expansion-of-tregs-and-suppression-of-th17-cells
#1
Zhen Nan, Heng Fan, Qing Tang, Man Zhang, Meng Xu, Qianyun Chen, Yujin Liu, Yalan Dong, Hui Wu, Shuangjiao Deng
Bone marrow-derived mesenchymal stem cells (BMSCs) hold great promise for the treatment of inflammatory bowel disease (IBD) owing to the immunosuppressive property and tissue healing potential. The balance of Treg/Th17 plays a crucial part in BMSC-mediated immunosuppression. Interleukin (IL)-35 is a newly identified anti-inflammatory cytokine required for the expansion of regulatory T cells (Tregs) and suppression of Th17 cells differentiation. It could amplify the immunosuppressive property of BMSCs by transfecting into BMSCs...
March 7, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29233896/retargeting-lentiviruses-via-spycatcher-spytag-chemistry-for-gene-delivery-into-specific-cell-types
#2
Nagarjun Kasaraneni, Ana M Chamoun-Emanuelli, Gus Wright, Zhilei Chen
We report a simple strategy for the creation of lentiviral vectors specific to any desired target cells. SpyTag is inserted into an engineered Sindbis virus envelope protein and displayed on the lentivirus surface to create Sindbis virus-SpyTag pseudoparticles (Sind-SpyTag-pp). The SpyTag serves as the covalent anchoring site for a target-cell-specific cell-binding protein (CBP) that is fused to a truncated SpyCatcher (SpyCatcherΔ). Target-cell-specific lentiviruses are created by mixing the Sind-SpyTag-pp and CBP-SpyCatcherΔ in vitro We first used a HER2-binding designed ankyrin repeat protein (DARPin...
December 12, 2017: MBio
https://www.readbyqxmd.com/read/29196603/effects-of-chondroitin-sulfate-proteoglycan-4-ng2-cspg4-on-soft-tissue-sarcoma-growth-depend-on-tumor-developmental-stage
#3
Shu-Hsuan Claire Hsu, Puviindran Nadesan, Vijitha Puviindran, William B Stallcup, David G Kirsch, Benjamin A Alman
Sarcomas, and the mesenchymal precursor cells from which they arise, express chondroitin sulfate proteoglycan 4 (NG2/CSPG4). However, NG2/CSPG4's function and its capacity to serve as a therapeutic target in this tumor type are unknown. Here, we used cells from human tumors and a genetically engineered autochthonous mouse model of soft-tissue sarcomas (STSs) to determine NG2/CSPG4's role in STS initiation and growth. Inhibiting NG2/CSPG4 expression in established murine and human STSs decreased tumor volume by almost two-thirds and cell proliferation rate by 50%...
February 16, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29150004/dynamics-of-indel-profiles-induced-by-various-crispr-cas9-delivery-methods
#4
Michael Kosicki, Sandeep S Rajan, Flaminia C Lorenzetti, Hans H Wandall, Yoshiki Narimatsu, Emmanouil Metzakopian, Eric P Bennett
The introduction of CRISPR/Cas9 gene editing in mammalian cells is a scientific breakthrough, which has greatly affected basic research and gene therapy. The simplicity and general access to CRISPR/Cas9 reagents has in an unprecedented manner "democratized" gene targeting in biomedical research, enabling genetic engineering of any gene in any cell, tissue, organ, and organism. The ability for fast, precise, and efficient profiling of the double-stranded break induced insertions and deletions (indels), mediated by any of the available programmable nucleases, is paramount to any given gene targeting approach...
2017: Progress in Molecular Biology and Translational Science
https://www.readbyqxmd.com/read/29016807/a-recombinant-lentiviral-pdgf-driven-mouse-model-of-proneural-glioblastoma
#5
Gilbert J Rahme, Bryan W Luikart, Chao Cheng, Mark A Israel
Background: Mouse models of glioblastoma (GBM), the most aggressive primary brain tumor, are critical for understanding GBM pathology and can contribute to the preclinical evaluation of therapeutic agents. Platelet-derived growth factor (PDGF) signaling has been implicated in the development and pathogenesis of GBM, specifically the proneural subtype. Although multiple mouse models of PDGF-driven glioma have been described, they require transgenic mice engineered to activate PDGF signaling and/or impair tumor suppressor genes and typically represent lower-grade glioma...
July 7, 2017: Neuro-oncology
https://www.readbyqxmd.com/read/28941274/advancements-in-the-design-and-scalable-production-of-viral-gene-transfer-vectors
#6
REVIEW
David Sharon, Amine Kamen
The last 10 years have seen a rapid expansion in the use of viral gene transfer vectors, with approved therapies and late stage clinical trials underway for the treatment of genetic disorders, and multiple forms of cancer, as well as prevention of infectious diseases through vaccination. With this increased interest and widespread adoption of viral vectors by clinicians and biopharmaceutical industries, there is an imperative to engineer safer and more efficacious vectors, and develop robust, scalable and cost-effective production platforms for industrialization...
September 23, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28881646/modeling-african-american-prostate-adenocarcinoma-by-inducing-defined-genetic-alterations-in-organoids
#7
Kenji Unno, Meejeon Roh, Young A Yoo, Yousef Al-Shraideh, Lu Wang, Larisa Nonn, Sarki A Abdulkadir
Genomic studies are rapidly identifying genetic alterations in human cancer, but functional validation of such alterations has been slow. Here, using human prostate cancer as a model, we have assessed the feasibility of engineering defined genetic alterations in well-known cancer driver genes to transform benign prostate epithelial organoids derived from African American men. Benign human prostate organoids were transduced with lentiviruses expressing MYC, shPTEN, shTP53 and AR, alone and in various combinations, to recapitulate prostate cancer development...
August 1, 2017: Oncotarget
https://www.readbyqxmd.com/read/28472770/modeling-african-american-prostate-adenocarcinoma-by-inducing-defined-genetic-alterations-in-organoids
#8
Kenji Unno, Meejeon Roh, Young A Yoo, Yousef Al-Shraideh, Lu Wang, Larisa Nonn, Sarki A Abdulkadir
Genomic studies are rapidly identifying genetic alterations in human cancer, but functional validation of such alterations has been slow. Here, using human prostate cancer as a model, we have assessed the feasibility of engineering defined genetic alterations in well-known cancer driver genes to transform benign prostate epithelial organoids derived from African American men. Benign human prostate organoids were transduced with lentiviruses expressing MYC, shPTEN, shTP53 and AR, alone and in various combinations, to recapitulate prostate cancer development...
April 19, 2017: Oncotarget
https://www.readbyqxmd.com/read/28062213/anti-fibrotic-effects-of-bone-morphogenetic-protein-7-modified-bone-marrow-mesenchymal-stem-cells-on-silica-induced-pulmonary-fibrosis
#9
Xiaoli Li, Guoliang An, Yan Wang, Di Liang, Zhonghui Zhu, Ximeng Lian, Piye Niu, Caixia Guo, Lin Tian
Silicosis is an occupational lung disease caused by exposure to small particles of crystalline silica, which ultimately results in diffuse pulmonary fibrosis. Evidence indicates an anti-fibrotic role of bone morphogenetic protein-7 (BMP-7) and bone marrow mesenchymal stem cells (BMSCs) in lung diseases. Therefore, strategies incorporating genetic engineering and stem cell biology might have a tremendous potential to treat critical injuries and diseases. Therefore, we modified BMSCs to overexpress the BMP-7 gene (BMP-7-BMSCs) by lentivirus transduction, and then evaluated whether fibrotic processes were inhibited by these cells in vivo...
February 2017: Experimental and Molecular Pathology
https://www.readbyqxmd.com/read/27934604/generation-of-induced-pluripotent-stem-cells-ipscs-stably-expressing-crispr-based-synergistic-activation-mediator-sam
#10
Kai Xiong, Yan Zhou, Poul Hyttel, Lars Bolund, Kristine Karla Freude, Yonglun Luo
Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated...
November 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27687658/-construction-of-a-lentivirus-vector-expressing-wheat-germ-agglutinin-and-its-infection-in-human-adipose-derived-stem-cells
#11
Juan Wang, Lian-Guo Zhang, Ning Li, Ming Lei, Yun Ma, Yuan-Hong Peng, Wei-Hua Bian
OBJECTIVE: To construct a lentivirus vector carrying wheat germ agglutinin (WGA) and evaluate its ability of tracing WGA in the brain of mice with ischemic brain injury. METHODS: WGA gene was inserted into the lentiviral vector Plvx IRES-ZsGreen1 using genetic engineering methods. 293T cells were transfected with the vector and 3 packaging plasmids (RPEV, PRRE, and VSVG) to obtain the recombinant lentivirus for infection of human adipose-derived stem cells (hADSCs)...
August 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
https://www.readbyqxmd.com/read/27574909/a-review-of-genetic-engineering-biotechnologies-for-enhanced-chronic-wound-healing
#12
John W Sessions, David G Armstrong, Sandra Hope, Brian D Jensen
Traditional methods for addressing chronic wounds focus on correcting dysfunction by controlling extracellular elements. This review highlights technologies that take a different approach - enhancing chronic wound healing by genetic modification to wound beds. Featured cutaneous transduction/transfection methods include viral modalities (i.e. adenoviruses, adeno-associated viruses, retroviruses, and lentiviruses) and conventional non-viral modalities (i.e. naked DNA injections, micro-seeding, liposomal reagents, particle bombardment, and electroporation)...
August 30, 2016: Experimental Dermatology
https://www.readbyqxmd.com/read/27404517/stem-cell-based-engineered-immunity-against-hiv-infection-in-the-humanized-mouse-model
#13
Anjie Zhen, Valerie Rezek, Cindy Youn, Jonathan Rick, Brianna Lam, Nelson Chang, Jerome Zack, Masakazu Kamata, Scott Kitchen
With the rapid development of stem cell-based gene therapies against HIV, there is pressing requirement for an animal model to study the hematopoietic differentiation and immune function of the genetically modified cells. The humanized Bone-marrow/Liver/Thymus (BLT) mouse model allows for full reconstitution of a human immune system in the periphery, which includes T cells, B cells, NK cells and monocytes. The human thymic implant also allows for thymic selection of T cells in autologous thymic tissue. In addition to the study of HIV infection, the model stands as a powerful tool to study differentiation, development and functionality of cells derived from hematopoietic stem cells (HSCs)...
July 2, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27357543/construction-of-an-anticancer-fusion-peptide-acfp-derived-from-milk-proteins-and-an-assay-of-anti-ovarian-cancer-cells-in-vitro
#14
Juan Zhou, Xue Yang, Wenxiao Zhang, Jing Wang, Cai Wei, Fang Gu, Ting Lei, Yide Qin
BACKGROUND: Some bioactive peptides derived from natural resources or synthesized by rational design have been shown to have very good anticancer effects. We designed an anticancer fusion peptide (ACFP) based on the structure of bovine lactoferricin (LfcinB) and hexapeptide (PGPIPN) derived from bovine milk protein. OBJECTIVE: To prepare ACFP through genetic engineering and study its antiovarian cancer activity. METHOD: ACFP gene was produced by a flexible link arm connecting LfcinB and PGPIPN...
2017: Anti-cancer Agents in Medicinal Chemistry
https://www.readbyqxmd.com/read/27333595/paralleled-comparison-of-vectors-for-the-generation-of-car-t-cells
#15
REVIEW
Di-Yuan Qin, Yong Huang, Dan Li, Yong-Sheng Wang, Wei Wang, Yu-Quan Wei
T-lymphocytes genetically engineered with the chimeric antigen receptor (CAR-T) have shown great therapeutic potential in cancer treatment. A variety of preclinical researches and clinical trials of CAR-T therapy have been carried out to lay the foundation for future clinical application. In these researches, several gene-transfer methods were used to deliver CARs or other genes into T-lymphocytes, equipping CAR-modified T cells with a property of recognizing and attacking antigen-expressing tumor cells in a major histocompatibility complex-independent manner...
September 2016: Anti-cancer Drugs
https://www.readbyqxmd.com/read/27155732/new-insights-and-current-tools-for-genetically-engineered-ge-sheep-and-goats
#16
REVIEW
A Menchaca, I Anegon, C B A Whitelaw, H Baldassarre, M Crispo
Genetically engineered sheep and goats represent useful models applied to proof of concepts, large-scale production of novel products or processes, and improvement of animal traits, which is of interest in biomedicine, biopharma, and livestock. This disruptive biotechnology arose in the 80s by injecting DNA fragments into the pronucleus of zygote-staged embryos. Pronuclear microinjection set the transgenic concept into people's mind but was characterized by inefficient and often frustrating results mostly because of uncontrolled and/or random integration and unpredictable transgene expression...
July 1, 2016: Theriogenology
https://www.readbyqxmd.com/read/27044116/prostate-epithelial-cell-of-origin-determines-cancer-differentiation-state-in-an-organoid-transformation-assay
#17
Jung Wook Park, John K Lee, John W Phillips, Patrick Huang, Donghui Cheng, Jiaoti Huang, Owen N Witte
The cell of origin for prostate cancer remains a subject of debate. Genetically engineered mouse models have demonstrated that both basal and luminal cells can serve as cells of origin for prostate cancer. Using a human prostate regeneration and transformation assay, our group previously demonstrated that basal cells can serve as efficient targets for transformation. Recently, a subpopulation of multipotent human luminal cells defined by CD26 expression that retains progenitor activity in a defined organoid culture was identified...
April 19, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/26819310/chimeric-filoviruses-for-identification-and-characterization-of-monoclonal-antibodies
#18
Philipp A Ilinykh, Xiaoli Shen, Andrew I Flyak, Natalia Kuzmina, Thomas G Ksiazek, James E Crowe, Alexander Bukreyev
UNLABELLED: Recent experiments suggest that some glycoprotein (GP)-specific monoclonal antibodies (MAbs) can protect experimental animals against the filovirus Ebola virus (EBOV). There is a need for isolation of MAbs capable of neutralizing multiple filoviruses. Antibody neutralization assays for filoviruses frequently use surrogate systems such as the rhabdovirus vesicular stomatitis Indiana virus (VSV), lentiviruses or gammaretroviruses with their envelope proteins replaced with EBOV GP or pseudotyped with EBOV GP...
April 2016: Journal of Virology
https://www.readbyqxmd.com/read/26808898/autism-like-behaviours-and-germline-transmission-in-transgenic-monkeys-overexpressing-mecp2
#19
Zhen Liu, Xiao Li, Jun-Tao Zhang, Yi-Jun Cai, Tian-Lin Cheng, Cheng Cheng, Yan Wang, Chen-Chen Zhang, Yan-Hong Nie, Zhi-Fang Chen, Wen-Jie Bian, Ling Zhang, Jianqiu Xiao, Bin Lu, Yue-Fang Zhang, Xiao-Di Zhang, Xiao Sang, Jia-Jia Wu, Xiu Xu, Zhi-Qi Xiong, Feng Zhang, Xiang Yu, Neng Gong, Wen-Hao Zhou, Qiang Sun, Zilong Qiu
Methyl-CpG binding protein 2 (MeCP2) has crucial roles in transcriptional regulation and microRNA processing. Mutations in the MECP2 gene are found in 90% of patients with Rett syndrome, a severe developmental disorder with autistic phenotypes. Duplications of MECP2-containing genomic segments cause the MECP2 duplication syndrome, which shares core symptoms with autism spectrum disorders. Although Mecp2-null mice recapitulate most developmental and behavioural defects seen in patients with Rett syndrome, it has been difficult to identify autism-like behaviours in the mouse model of MeCP2 overexpression...
February 4, 2016: Nature
https://www.readbyqxmd.com/read/26715207/primary-like-human-hepatocytes-genetically-engineered-to-obtain-proliferation-competence-display-hepatic-differentiation-characteristics-in-monolayer-and-organotypical-spheroid-cultures
#20
Natalie Herzog, Max Hansen, Sebastian Miethbauer, Kai-Uwe Schmidtke, Ursula Anderer, Amelie Lupp, Sebastian Sperling, Daniel Seehofer, Georg Damm, Katrin Scheibner, Jan-Heiner Küpper
Primary human hepatocytes are in great demand during drug development and in hepatology. However, both scarcity of tissue supply and donor variability of primary cells create a need for the development of alternative hepatocyte systems. By using a lentivirus vector system to transfer coding sequences of Upcyte® proliferation genes, we generated non-transformed stable hepatocyte cultures from human liver tissue samples. Here, we show data on newly generated proliferation-competent HepaFH3 cells investigated as conventional two-dimensional monolayer and as organotypical three-dimensional (3D) spheroid culture...
March 2016: Cell Biology International
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