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lentivirus genetic engineering

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
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
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)...
2016: Journal of Visualized Experiments: JoVE
Yide Qin, Juan Zhou, Wenxiao Zhang, Xue Yang, Jing Wang, Cai Wei, Fang Gu, Ting Lei
Some bioactive peptides derived from natural resources or synthesized by rational design have been shown to have very good anticancer effects. We designed and prepared an anticancer fusion peptide (ACFP) through genetic engineering and studied its antiovarian cancer activity by cell culture and lentivirus infection. Bovine lactoferricin (LfcinB) and hexapeptide (PGPIPN) were connected by a flexible link arm (GGGGS) to produce an ACFP gene. ACFP was inductively expressed in Escherichia coli by the recombinant plasmid pGEX-KG-ACFP...
June 26, 2016: Anti-cancer Agents in Medicinal Chemistry
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
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
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
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
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
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
Zepeng Qu, Yuan Guan, Lu Cui, Jian Song, Junjie Gu, Hanzhi Zhao, Lei Xu, Lixia Lu, Ying Jin, Guo-Tong Xu
INTRODUCTION: Degenerative retinal diseases like age-related macular degeneration (AMD) are the leading cause of blindness. Cell transplantation showed promising therapeutic effect for such diseases, and embryonic stem cell (ESC) is one of the sources of such donor cells. Here, we aimed to generate retinal progenitor cells (RPCs) from rat ESCs (rESCs) and to test their therapeutic effects in rat model. METHODS: The rESCs (DA8-16) were cultured in N2B27 medium with 2i, and differentiated to two types of RPCs following the SFEBq method with modifications...
2015: Stem Cell Research & Therapy
Y J Xu, W R Chen, D P Li, L X Song, J Q Wu, P Zhang, Z Y Li, Y H Huang
We determined whether genetically engineered immature dendritic cells (imDCs) mediated by lentiviral vectors alleviate acute graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo-BMT) in mice. We introduced the mouse chemokine receptor 7 (Ccr7) gene into the bone marrow-derived imDCs of C57BL/6 mice to construct genetically engineered imDCs. A 1:1 mixture of bone marrow and spleen cells from the donors was injected into the recipients, which were divided into four groups: radiation, transplantation, empty vector, and transgenic imDC groups...
2015: Genetics and Molecular Research: GMR
Xiaoyu Chen, Manuel A F V Gonçalves
Genome editing based on sequence-specific designer nucleases, also known as programmable nucleases, seeks to modify in a targeted and precise manner the genetic information content of living cells. Delivering into cells designer nucleases alone or together with donor DNA templates, which serve as surrogate homologous recombination (HR) substrates, can result in gene knockouts or gene knock-ins, respectively. As engineered replication-defective viruses, viral vectors are having an increasingly important role as delivery vehicles for donor DNA templates and designer nucleases, namely, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated Cas9 (CRISPR-Cas9) nucleases, also known as RNA-guided nucleases (RGNs)...
March 2016: Molecular Therapy: the Journal of the American Society of Gene Therapy
Constant Morez, Michela Noseda, Marta Abreu Paiva, Elisa Belian, Michael D Schneider, Molly M Stevens
Generation of de novo cardiomyocytes through viral over-expression of key transcription factors represents a highly promising strategy for cardiac muscle tissue regeneration. Although the feasibility of cell reprogramming has been proven possible both in vitro and in vivo, the efficiency of the process remains extremely low. Here, we report a chemical-free technique in which topographical cues, more specifically parallel microgrooves, enhance the directed differentiation of cardiac progenitors into cardiomyocyte-like cells...
November 2015: Biomaterials
Gang Xu, Yanwu Guo, Zhiyuan Seng, Gang Cui, Jianqiang Qu
Bone marrow-derived mesenchymal stem cells (BMSCs) are promising gene vehicles for cancer gene therapy. In our previous study, we reported that BMSCs expressing interleukin (IL)-18 effectively inhibit the growth of glioma in rats. In the present study, we further detected the effect of BMSCs co-expressing IL-18 and interferon (IFN)-β, both of which are immunostimulatory cytokines. BMSCs were genetically engineered to express IL-18 and IFN-β by transfection of recombinant lentivirus-mediated gene transfer...
October 2015: Oncology Reports
Zhen Wang, Da-chuan Liang, Jie-yu Bai, Ning Kang, Jun-yu Feng, Zi-quan Yang
OBJECTIVE: To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo. METHODS: Rabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment...
May 2015: Zhongguo Gu Shang, China Journal of Orthopaedics and Traumatology
Shin-Heng Chiou, Ian P Winters, Jing Wang, Santiago Naranjo, Crissy Dudgeon, Fiona B Tamburini, Jennifer J Brady, Dian Yang, Barbara M Grüner, Chen-Hua Chuang, Deborah R Caswell, Hong Zeng, Pauline Chu, Grace E Kim, Darren R Carpizo, Seung K Kim, Monte M Winslow
Pancreatic ductal adenocarcinoma (PDAC) is a genomically diverse, prevalent, and almost invariably fatal malignancy. Although conventional genetically engineered mouse models of human PDAC have been instrumental in understanding pancreatic cancer development, these models are much too labor-intensive, expensive, and slow to perform the extensive molecular analyses needed to adequately understand this disease. Here we demonstrate that retrograde pancreatic ductal injection of either adenoviral-Cre or lentiviral-Cre vectors allows titratable initiation of pancreatic neoplasias that progress into invasive and metastatic PDAC...
July 15, 2015: Genes & Development
Wanqiu Chen, David J Baylink, Justin Brier-Jones, Amanda Neises, Jason B Kiroyan, Charles H Rundle, Kin-Hing William Lau, Xiao-Bing Zhang
Substantial advances have been made in the past two decades in the management of osteoporosis. However, none of the current medications can eliminate the risk of fracture and rejuvenate the skeleton. To this end, we recently reported that transplantation of hematopoietic stem/progenitor cells (HSCs) or Sca1(+) cells engineered to overexpress FGF2 results in a significant increase in lamellar bone matrix formation at the endosteum; but this increase was attended by the development of secondary hyperparathyroidism and severe osteomalacia...
July 21, 2015: Proceedings of the National Academy of Sciences of the United States of America
Ana M Chamoun-Emanuelli, Gus Wright, Smith Roger, Robert C Münch, Christian J Buchholz, Zhilei Chen
Gene therapy represents a promising therapeutic paradigm for addressing many disorders, but the absence of a vector that can be robustly and reproducibly functionalized with cell-homing functionality to mediate the delivery of genetic cargo specifically to target cells following systemic administration has stood as a major impediment. In this study, a high-affinity protein-protein pair comprising a splicing-deficient naturally split intein was used as molecular Velcro to append a HER2/neu-binding protein (DARPin) onto the surface of a binding-deficient, fusion-competent lentivirus...
December 2015: Biotechnology and Bioengineering
Parwiz Abrahimi, William G Chang, Martin S Kluger, Yibing Qyang, George Tellides, W Mark Saltzman, Jordan S Pober
RATIONALE: The participation of endothelial cells (EC) in many physiological and pathological processes is widely modeled using human EC cultures, but genetic manipulation of these untransformed cells has been technically challenging. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) technology offers a promising new approach. However, mutagenized cultured cells require cloning to yield homogeneous populations, and the limited replicative lifespan of well-differentiated human EC presents a barrier for doing so...
July 3, 2015: Circulation Research
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