scholarly journals Effects of transgene expression level per cell in mice livers on induction of transgene-specific immune responses after hydrodynamic gene transfer

Gene Therapy ◽  
2016 ◽  
Vol 23 (7) ◽  
pp. 565-571 ◽  
Author(s):  
Y Yin ◽  
Y Takahashi ◽  
A Hamana ◽  
M Nishikawa ◽  
Y Takakura
Keyword(s):  
Gene Transfer ◽  
Immune Responses ◽  
Transgene Expression ◽  
Expression Level ◽  
Transgene Expression Level

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 Increased transgene expression level of rabies virus vector for transsynaptic tracing

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0180960 ◽  
Author(s):  
Shinya Ohara ◽  
Yasuhiro Sota ◽  
Sho Sato ◽  
Ken-Ichiro Tsutsui ◽  
Toshio Iijima
Keyword(s):  
Rabies Virus ◽  
Transgene Expression ◽  
Virus Vector ◽  
Expression Level ◽  
Transsynaptic Tracing ◽  
Transgene Expression Level

scholarly journals High transgene expression is associated with systemic GFP silencing in Nicotiana benthamiana

2021 ◽  
Author(s):  
Bill Hendrix ◽  
Paul Hoffer ◽  
Rick Sanders ◽  
Steve Schwartz ◽  
Wei Zheng ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Transgene Expression ◽  
Fluorescent Protein ◽  
Integration Site ◽  
Crop Protection ◽  
Transgene Silencing ◽  
Expression Level ◽  
Transgenic Lines ◽  
Transgene Expression Level

AbstractGene silencing in plants using topical dsRNA is a new approach that has the potential to be a sustainable component of the agricultural production systems of the future. However, more research is needed to enable this technology as an economical and efficacious supplement to current crop protection practices. Systemic gene silencing is one key enabling aspect. The objective of this research was to better understand systemic transgene silencing in Nicotiana benthamiana. Previous reports details sequencing of the integration site of the Green Fluorescent Protein (GFP) transgene in the well-known N. benthamiana GFP16C event revealed inadvertent co-integration of part of a bacterial transposase. To determine the effect of this transgene configuration on systemic silencing, new GFP transgenic lines with or without the transposase sequences were produced. GFP expression levels in the 19 single-copy events and three hemizygous 16C lines produced for this study ranged from 50-72% of the homozygous 16C line. GFP expression was equivalent to 16C in a two-copy event. Local GFP silencing was observed in all transgenic and 16C hemizygous lines after topical application of delivery formulations with a GFP targeting dsRNA. The 16C-like systemic silencing phenotype was only observed in the two-copy line. The partial transposase had no impact on transgene expression level, local GFP silencing, small RNA abundance and distribution, or systemic GFP silencing in the transgenic lines. We conclude that high transgene expression level is a key enabler of systemic transgene silencing in N. benthamiana.

scholarly journals Rabies Virus Vector Transgene Expression Level and Cytotoxicity Improvement Induced by Deletion of Glycoprotein Gene

PLoS ONE ◽  
2013 ◽  
Vol 8 (11) ◽  
pp. e80245 ◽  
Author(s):  
Shinya Ohara ◽  
Sho Sato ◽  
Kei Oyama ◽  
Ken-Ichiro Tsutsui ◽  
Toshio Iijima
Keyword(s):  
Rabies Virus ◽  
Transgene Expression ◽  
Virus Vector ◽  
Expression Level ◽  
Glycoprotein Gene ◽  
Transgene Expression Level

Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice

Blood ◽  
2004 ◽  
Vol 103 (10) ◽  
pp. 3700-3709 ◽  
Author(s):  
Antonia Follenzi ◽  
Manuela Battaglia ◽  
Angelo Lombardo ◽  
Andrea Annoni ◽  
Maria Grazia Roncarolo ◽  
...  
Keyword(s):  
T Cells ◽  
Gene Transfer ◽  
Immune Responses ◽  
Transgene Expression ◽  
Factor Ix ◽  
Stable Gene ◽  
Human Factor Ix ◽  
Immunocompetent Mice

Abstract Stable gene replacement by in vivo administration of lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immunocompetent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgene expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for the gene therapy of hemophilias and other liver-based diseases.

scholarly journals Systemic GFP silencing is associated with high transgene expression in Nicotiana benthamiana

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0245422
Author(s):  
Bill Hendrix ◽  
Paul Hoffer ◽  
Rick Sanders ◽  
Steve Schwartz ◽  
Wei Zheng ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Transgene Expression ◽  
Fluorescent Protein ◽  
Integration Site ◽  
Crop Protection ◽  
Transgene Silencing ◽  
Expression Level ◽  
Transgenic Lines ◽  
Transgene Expression Level

Gene silencing in plants using topical dsRNA is a new approach that has the potential to be a sustainable component of the agricultural production systems of the future. However, more research is needed to enable this technology as an economical and efficacious supplement to current crop protection practices. Systemic gene silencing is one key enabling aspect. The objective of this research was to better understand topically-induced, systemic transgene silencing in Nicotiana benthamiana. A previous report details sequencing of the integration site of the Green Fluorescent Protein (GFP) transgene in the well-known N. benthamiana GFP16C event. This investigation revealed an inadvertent co-integration of part of a bacterial transposase in this line. To determine the effect of this transgene configuration on systemic silencing, new GFP transgenic lines with or without the transposase sequences were produced. GFP expression levels in the 19 single-copy events and three hemizygous GFP16C lines produced for this study ranged from 50–72% of the homozygous GFP16C line. GFP expression was equivalent to GFP16C in a two-copy event. Local GFP silencing was observed in all transgenic and GFP16C hemizygous lines after topical application of carbon dot-based formulations containing a GFP targeting dsRNA. The GFP16C-like systemic silencing phenotype was only observed in the two-copy line. The partial transposase had no impact on transgene expression level, local GFP silencing, small RNA abundance and distribution, or systemic GFP silencing in the transgenic lines. We conclude that high transgene expression level is a key enabler of topically-induced, systemic transgene silencing in N. benthamiana.

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