scholarly journals Optimized Lentiviral Vector Design Improves Titer and Transgene Expression of Vectors Containing the Chicken β-Globin Locus HS4 Insulator Element

2009 ◽  
Vol 17 (4) ◽  
pp. 667-674 ◽  
Author(s):  
Hideki Hanawa ◽  
Motoko Yamamoto ◽  
Huifen Zhao ◽  
Takashi Shimada ◽  
Derek A Persons
Keyword(s):  
Transgene Expression ◽  
Lentiviral Vector ◽  
Insulator Element

scholarly journals Retrograde Transgene Expression via Neuron-Specific Lentiviral Vector Depends on Both Species and Input Projections

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1387
Author(s):  
Yukiko Otsuka ◽  
Hitomi Tsuge ◽  
Shiori Uezono ◽  
Soshi Tanabe ◽  
Maki Fujiwara ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Immune Responses ◽  
Envelope Glycoprotein ◽  
Transgene Expression ◽  
Histological Analysis ◽  
Lentiviral Vector ◽  
Cortical Input ◽  
Input System ◽  
Vesicular Stomatitis ◽  
Retrograde Gene Transfer

For achieving retrograde gene transfer, we have so far developed two types of lentiviral vectors pseudotyped with fusion envelope glycoprotein, termed HiRet vector and NeuRet vector, consisting of distinct combinations of rabies virus and vesicular stomatitis virus glycoproteins. In the present study, we compared the patterns of retrograde transgene expression for the HiRet vs. NeuRet vectors by testing the cortical input system. These vectors were injected into the motor cortex in rats, marmosets, and macaques, and the distributions of retrograde labels were investigated in the cortex and thalamus. Our histological analysis revealed that the NeuRet vector generally exhibits a higher efficiency of retrograde gene transfer than the HiRet vector, though its capacity of retrograde transgene expression in the macaque brain is unexpectedly low, especially in terms of the intracortical connections, as compared to the rat and marmoset brains. It was also demonstrated that the NeuRet but not the HiRet vector displays sufficiently high neuron specificity and causes no marked inflammatory/immune responses at the vector injection sites in the primate (marmoset and macaque) brains. The present results indicate that the retrograde transgene efficiency of the NeuRet vector varies depending not only on the species but also on the input projections.

scholarly journals Stability of Lentiviral Vector-Mediated Transgene Expression in the Brain in the Presence of Systemic Antivector Immune Responses

2005 ◽  
Vol 16 (6) ◽  
pp. 741-751 ◽  
Author(s):  
Evelyn Abordo-Adesida ◽  
Antonia Follenzi ◽  
Carlos Barcia ◽  
Sandra Sciascia ◽  
Maria G. Castro ◽  
...  
Keyword(s):  
Immune Responses ◽  
Transgene Expression ◽  
Lentiviral Vector ◽  
The Brain

scholarly journals Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts

2019 ◽  
Vol 22 (8) ◽  
pp. 1020-1025 ◽  
Author(s):  
V. R. Beklemisheva ◽  
A. G. Menzorov
Keyword(s):  
Transgene Expression ◽  
Viral Vectors ◽  
Lentiviral Vector ◽  
Sendai Virus ◽  
Homo Sapiens ◽  
Fluorescent Proteins ◽  
American Mink ◽  
Ips Cells ◽  
Canis Lupus Familiaris ◽  
Induced Pluripotent

Generation of induced pluripotent stem (iPS) cells expanded possibilities of pluripotency and early development studies. Generation of order Carnivora iPS cells from dog (Canis lupus familiaris), snow leopard (Panthera uncia), and American mink (Neovison vison) was previously reported. The aim of the current study was to examine conditions of pinniped fbroblast reprogramming. Pinnipeds are representatives of the suborder Caniformia sharing conservative genomes. There are several ways to deliver reprogramming transcription factors: RNA, proteins, plasmids, viral vectors etc. The most effective delivery systems for mouse and human cells are based on viral vectors. We compared a lentiviral vector which integrates into the genome and a Sendai virus­based vector, CytoTune EmGFP Sendai Fluorescence Reporter. The main advantage of Sendai virus­based vectors is that they do not integrate into the genome. We performed delivery of genetic constructions carrying fluorescent proteins to fbroblasts of seven Pinnipeds: northern fur seal (Callorhinus ursinus), Steller sea lion (Eumetopias jubatus), walrus (Odobenus rosmarus), bearded seal (Erignathus barbatus), Baikal seal (Pusa sibirica), ringed seal (Phoca hispida), and spotted seal (Phoca largha). We also transduced American mink (N. vison), human (Homo sapiens), and mouse (Mus musculus) fbroblasts as a control. We showed that the Sendai virus­based transduction system provides transgene expression one­two orders of magnitude higher than the lentiviral system at a comparable multiplicity of infection. Also, transgene expression after Sendai virus­based transduction is quite stable and changes only slightly at day four compared to day two. These data allow us to suggest that Sendai virus­based vectors are preferable for generation of Pinniped iPS cells.

scholarly journals Host Cell Detection of Noncoding Stuffer DNA Contained in Helper-Dependent Adenovirus Vectors Leads to Epigenetic Repression of Transgene Expression

2009 ◽  
Vol 83 (17) ◽  
pp. 8409-8417 ◽  
Author(s):  
P. Joel Ross ◽  
Michael A. Kennedy ◽  
Robin J. Parks
Keyword(s):  
Transgene Expression ◽  
Great Promise ◽  
Cell Detection ◽  
Nuclear Factors ◽  
Insulator Element ◽  
Pml Bodies ◽  
Eukaryotic Dna ◽  
Foreign Dna ◽  
Mechanistic Basis ◽  
Epigenetic Repression

ABSTRACT Helper-dependent adenovirus (hdAd) vectors have shown great promise as therapeutic gene delivery vehicles in gene therapy applications. However, the level and duration of gene expression from hdAd can differ considerably depending on the nature of the noncoding stuffer DNA contained within the vector. For example, an hdAd containing 22 kb of prokaryotic DNA (hdAd-prok) expresses its transgene 60-fold less efficiently than a similar vector containing eukaryotic DNA (hdAd-euk). Here we have determined the mechanistic basis of this phenomenon. Although neither vector was subjected to CpG methylation and both genomes associated with cellular histones to similar degrees, hdAd-prok chromatin was actively deacetylated. Insertion of an insulator element between the transgene and the bacterial DNA derepressed hdAd-prok, suggesting that foreign DNA nucleates repressive chromatin structures that spread to the transgene. We found that Sp100B/Sp100HMG and Daxx play a role in repressing transgene expression from hdAd and act independently of PML bodies. Thus, we have identified nuclear factors involved in recognizing foreign DNA and have determined the mechanism by which associated genes are repressed.

scholarly journals Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

Gene Therapy ◽  
2003 ◽  
Vol 10 (17) ◽  
pp. 1446-1457 ◽  
Author(s):  
Y Bai ◽  
Y Soda ◽  
K Izawa ◽  
T Tanabe ◽  
X Kang ◽  
...  
Keyword(s):  
Human Blood ◽  
Blood Cells ◽  
Transgene Expression ◽  
Lentiviral Vector ◽  
Third Generation ◽  
Human Blood Cells

Real-Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction as a Method for Determining Lentiviral Vector Titers and Measuring Transgene Expression

2003 ◽  
Vol 14 (6) ◽  
pp. 497-507 ◽  
Author(s):  
Gregory Lizée ◽  
Joeri L. Aerts ◽  
Monica I. Gonzales ◽  
Nachimuthu Chinnasamy ◽  
Richard A. Morgan ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Transgene Expression ◽  
Lentiviral Vector ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Toward Tightly Tuned Gene Expression Following Lentiviral Vector Transduction

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1427
Author(s):  
Audrey Page ◽  
Floriane Fusil ◽  
François-Loïc Cosset
Keyword(s):  
Gene Expression ◽  
Transgene Expression ◽  
Lentiviral Vector ◽  
Basic Research ◽  
Lentiviral Vectors ◽  
Retroviral Vectors ◽  
Clinical Applications ◽  
Long Terminal Repeats ◽  
Vector Systems ◽  
Gene Excision

Lentiviral vectors are versatile tools for gene delivery purposes. While in the earlier versions of retroviral vectors, transgene expression was controlled by the long terminal repeats (LTRs), the latter generations of vectors, including those derived from lentiviruses, incorporate internal constitutive or regulated promoters in order to regulate transgene expression. This allows to temporally and/or quantitatively control transgene expression, which is required for many applications such as for clinical applications, when transgene expression is required in specific tissues and at a specific timing. Here we review the main systems that have been developed for transgene regulated expression following lentiviral gene transfer. First, the induction of gene expression can be triggered either by external or by internal cues. Indeed, these regulated vector systems may harbor promoters inducible by exogenous stimuli, such as small molecules (e.g., antibiotics) or temperature variations, offering the possibility to tune rapidly transgene expression in case of adverse events. Second, expression can be indirectly adjusted by playing on inserted sequence copies, for instance by gene excision. Finally, synthetic networks can be developed to sense specific endogenous signals and trigger defined responses after information processing. Regulatable lentiviral vectors (LV)-mediated transgene expression systems have been widely used in basic research to uncover gene functions or to temporally reprogram cells. Clinical applications are also under development to induce therapeutic molecule secretion or to implement safety switches. Such regulatable approaches are currently focusing much attention and will benefit from the development of other technologies in order to launch autonomously controlled systems.

Physiological regulation of transgene expression by a lentiviral vector containing the A2UCOE linked to a myeloid promoter

Gene Therapy ◽  
2011 ◽  
Vol 19 (10) ◽  
pp. 1018-1029 ◽  
Author(s):  
C Brendel ◽  
U Müller-Kuller ◽  
S Schultze-Strasser ◽  
S Stein ◽  
L Chen-Wichmann ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Lentiviral Vector ◽  
Physiological Regulation
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