Non-union occurring in structural bone grafting is a major problem in allograft transplantation because of impaired interaction between the host and graft tissue.
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Vectorology - GEG Tech top picks
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BigField GEG Tech's insight:
In this study, the scientists designed a recombinant lentiviral vector expressing four fragments of mouse CCR3 shRNA (pLVX‑mCCR3‑1+2+3+4‑shRNA). They show that downregulation of mCCR3 significantly inhibited proliferation of the eosinophils. Furthermore, the present study found that the downregulation of mCCR3 significantly promoted apoptosis of the eosinophils. These results suggest that mCCR3 silencing may serve as a putative approach for the treatment of allergic rhinitis. 
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The available different lentiviral vectors for conditional shRNA expression cassette delivery co-express additional genes that allow the use of fluorescent proteins for color-coded combinatorial RNAi or monitoring RNAi induction (pGLTR-FP), selection of transduced cells (pGLTR-S), and the generation of conditional cell lines using a one-vector system (pGLTR-X).
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In this work, the authors designed self-inactivating lentiviral vectors expressing multiple siRNAs targeting the CCR5 cellular gene as well as vif and tat/revviral transcripts, under the control of different RNA polymerase III promoters (U6, 7SK, H1) to inhibit different steps of virus life cycle. The efficacy of selected developed combinatorial vectors in interfering with viral replication was evaluated in human primary CD4+ T lymphocytes and the scientists observed two effective anti-HIV combinatorial vectors that conferred protection against R5- and X4- tropic viruses. These findings contribute to gain further insights in the design of RNAi-based gene therapy approaches against HIV-1 for clinical application.
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In this study, the scientists construct an RNA interference (RNAi) recombinant lentiviral vector particle targeting DEPDC7 in order to knockdown its gene expression in human hepatocellular carcinoma cell line HepG2. They observe that migration and invasion of transduced HepG2 cells were significantly increased. These data suggest that shRNA-mediated knockdown of DEPDC7 enable promotion of cell migration and invasion.
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In this study, the authors investigated the contribution of epigenetic factors HDAC1 and DAXX to the developmental perturbation hypothesized to underlie schizophrenia (SZ). For this purpose, they designed lentiviral vectors carrying short hairpin RNA interference (shRNAi) for HDAC1 and DAXX. Their results point to a key role of both HDAC1 and DAXX in the regulation of GAD67 in GABAergic HiB5 cells, strongly suggesting that these epigenetic/transcription factors contribute to mechanisms underlying GABA cell dysfunction in SZ.
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Here, the authors designed shRNA lentiviral vectors which target GFP and evaluated the effects of various factors on silencing efficiency including stem length, loop sequence, antisense location as well as the ratio between AGO2 and shRNA. They found that 19 ~ 21-bp stem and 6- or 9-nt loop structure in the sense-loop-antisense (S-L-AS) orientation was an optimal design in the AGO2-shRNA system. Furthermore, they observed that simultaneous expression of shRNA and AGO2 can achieve robust silencing effect. However, the titers of packaged lentivirus from constitutive expression of AGO2 vector were extremely low, severely limiting its broad application. Here, they demonstrated that the problem can be significantly improved by using the inducible expression of AGO2 lentiviral system.
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CTDP1 was overexpressed in many human lung cancer.In this work, the authors used a shRNA lentiviral vector to knockdown CTDP1 expression. They observe that inhibition of CTDP1 expression significantly suppressed cell growth, induced G0/G1 phase arrest and repressed cell colony formation. These data show that CTDP1 played an important role in cell proliferation and may be a useful therapeutic target in human lung cancer.
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RNAi is a powerful tool to achieve suppression of a specific gene expression. However, delivery and long-term expression of shRNAs should be carefully optimized for efficient knock down of target gene without causing cytotoxicity in mammalian cells. In this suty, the authors describe protocols for the development of shRNA against a major HIV co-receptor/chemokine receptor CCR5 and the use of lentiviral vectors for stable shRNA delivery and expression in human primary hematopoietic stem/progenitor cells.
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Here, the authors used recombinant lentiviral RNAi vectors for β1 or β2-ARs to study the functional roles of the β-ARs subtypes, β1- and β2- ARs, in the hippocampal cornu ammonis 3 (CA3) region. Their results suggested that the β1- and β2-ARs were involved in hippocampus dependent memory consolidation.
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The scientists investigated the effects of HSP27 expression on cytoskeleton dynamics and contractile function of human bladder smooth muscle cells (BSMCs) in vitro. In this goal, they used lentiviral vectors to overexpress or knock down HSP27. They show that the structure of actin filaments in HSP27 over-expressed cells recovered and F/G-actin ratio significantly increased at 12 h after stretching which is not the case in HSP27 knock-down cells. In addition, the contractile force of BSMCs was enhanced by over-expression of HSP27 and attenuated by knock-down of HSP27, suggesting a pivotal role of HSP27 in regulating bladder smooth muscle contraction.
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To investigate the role of SOCS3 in erythropoiesis, the author used a shRNA lentiviral vectors which knockdowns this gene. They found that the inhibition of the gene induced erythroid expansion in HSCs. Conversely, Ectopic expression of SOCS3 in progenitor cells blocked erythroid expansion and erythroid colony formation of HSCs. These results provided new insight into the mechanism of erythropoietic development.
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In this study, the authors used a lentiviral vector to overexpress miR-9 which is negatively regulates by N-myc. They show that miR-9 can inhibit the proliferation of granule cell precursors (GCPs) and highlight N-myc like a key switch regulating of this process.
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In this wotk, the scientists investigate the effect of silencing of hexokinase genes (HK1, HK2, and HK3) in colorectal cancer (HT-29, SW 480, HCT-15, RKO, and HCT 116) and melanoma (MDA-MB-435S and SK-MEL-28) cell lines using short hairpin RNA (shRNA) lentiviral vectors. They demonstrated that simultaneous inactivation of HK1 and HK2 was sufficient to decrease proliferation and viability of melanoma and colorectal cancer cells. Their results suggest that HK1 and HK2 could be the key therapeutic targets for reducing aerobic glycolysis in examined cancers.
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In this study, the scientists used a lentiviral vector which express shRNA Notch1, known as a cell fate regulator, to investigate its role in the mechanism of endometriosis. The results showed that LV-Notch1-shRNA led to a significant decline of clonogenicity and migration in human endometrial stem cells in vitro, as well as the size of endometriotic lesions in murine models. These data provide evidence that specific inhibition of Notch1 alters endometriotic tissue growth and progression, and may represent a promising potential therapeutic avenue.
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In this study, the authors use a lentiviral vectors expressing sh RNA which target the Toll-like receptor 4 (TLR4). They transduced human lens epithelial cell line HLEC and observed a decrease of proliferation and an extended cell cycle.
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Liver fibrosis, the accumulation of extracellular matrix proteins caused by chronic liver damage, results from the activation of hepatic stellate cells. In a new study, Arriazu et al. explore in detail the mechanisms by which the multifunctional protein osteopontin drives hepatic fibrosis, and show that interaction with HMGB1 mediates the fibrogenic response.
BigField GEG Tech's insight:
The authors identified RAGE as the key mediator of HMGB1 effects on HSCs by using lentiviral expression vectors to achieve small hairpin expression knockdown of either TLR2, TLR4 and TLR9 or RAGE, and then showed that only knockdown of RAGE decreased HMGB1-induced collagen expression.
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The aim of the present study was to clarify whether βcatenin was associated with the proliferation and progress of glioma. To do that, scientists have used lentiviral vectors expressing βcatenin short hairpin (sh)RNA. Their results demonstrated that knockdown of βcatenin expression significantly inhibited the progression of human glioma cancer cells, in vitro and in vivo; thus suggesting that βcatenin silencing may be a novel therapy for the treatment of human glioma.
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In this sutdy, the authors used lentiviral vectors to oiverexpressed or silenced (shRNA) ITGα5 and ITGβ1 which are fibronectin receptors that are abundantly expressed on the surface of vascular smooth muscle cells (VSMCs). ITGβ1 was markedly associated with the proliferation and migration of VSMCs whereas ITGα5 did not exert any effects on VSMCs. The results of the present study may provide a possible therapeutic target for the prevention and treatment of early vascular disease associated with VSMCs.
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Here, Tomas Bjorklund review design choices for enabling expression of two functional protein or RNA sequences from a single viral vector. These type of tools are very useful in neuroscience-related field of neuronal control and modulation, e.g., using optogenetics or DREADDs, but are also desirable in applications of CRISPR/Cas9 in situ genome editing and more refined therapeutic approaches.
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Here the authors investigate the effect of promoter methylation of cholesterol 25-hydroxylase (CH25H) gene. They observe that CH25H-knockdown by transductionof shRNA lentiviral vector into the cell lines partially protected the cells from DAC-induced cell death. Further investigations of the promoter analysis of CH25H gene and therapeutic effects of DNMT inhibitors on MDS/leukemia will be warranted.
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Pulmonary arterial hypertension (PAH) is a life-threatening disorder that ultimately causes heart failure. In this study, the authors used a shRNA lentiviral vector which targets Bcl2 and another which expresses hypoxia response element (HRE) to understand the causes of PAH. Their results suggest that HRE-mediated Bcl-2 inhibition can effectively attenuate hypoxia-induced apoptosis resistance in pulmonary microvascular endothelial cells by downregulating Bcl-2 expression and upregulating caspase-3 and P53 expression. This study therefore reveals critical insight into potential therapeutic targets for treating PAH.
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Here, the authors downregulation LAPTM4B gene expression in NIH:OVCAR3 cells thanks to a sh lentiviral vector. The transduced cells exhibited significant decreases in cell motility and invasion. Furthermore, LAPTM4B depletion resulted in a significant decrease in PCNA, VEGF, MMP2, MMP9 and CDK12 expression. These results suggest that LAPTM4B expression could be an oncogene-inducing feature of invasive ovarian cancer cells and could be a potential therapeutic target for ovarian cancer treatment.
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In this study, the authors investigate the effect of growth-associated protein-43 (GAP-43) on bone marrow mesenchymal stem cell (BMSC) differentiation in a rat model of traumatic optic neuropathy (TON). They realize GAP‑43 overexpression and GAP‑43 knockdown thanks different lentiviral vectors. In this way, they show that GAP‑43 promoted BMSC differentiation into neuron-like cells, and intravitreally injected GAP-43/BMSCs promoted the process of nerve repair in a rat model of TON.
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Through the use of a shRNA lentiviral vector which targets Irf6, the scientists show that is a mediator of TGFβ3, which regulates an epithelial mesenchymal transition and fusion process during the embryonic palate development.
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In this study, the scientists evaluated the effects of local knockdown of the TLR4 signaling pathway in a mouse segmental femoral graft model. Allografts were coated with freeze-dried lentiviral vectors that encoded TLR4 and myeloid differentiation primary response gene 88 (MyD88) short-hairpin RNA (shRNA), which were individually transplanted into the mice. Their results show that inhibition of TLR4 and MyD88 might reduce immune responses and ameliorate allograft rejection.