scholarly journals Viral Transduction of Human Rod Opsin or Channelrhodopsin Variants to Mouse ON Bipolar Cells Does Not Impact Retinal Anatomy or Cause Measurable Death in the Targeted Cells

2021 ◽  
Vol 22 (23) ◽  
pp. 13111
Author(s):  
Phillip Wright ◽  
Jessica Rodgers ◽  
Jonathan Wynne ◽  
Paul N. Bishop ◽  
Robert J. Lucas ◽  
...  

The viral gene delivery of optogenetic actuators to the surviving inner retina has been proposed as a strategy for restoring vision in advanced retinal degeneration. We investigated the safety of ectopic expression of human rod opsin (hRHO), and two channelrhodopsins (enhanced sensitivity CoChR-3M and red-shifted ReaChR) by viral gene delivery in ON bipolar cells of the mouse retina. Adult Grm6Cre mice were bred to be retinally degenerate or non-retinally degenerate (homozygous and heterozygous for the rd1Pde6b mutation, respectively) and intravitreally injected with recombinant adeno-associated virus AAV2/2(quad Y-F) serotype containing a double-floxed inverted transgene comprising one of the opsins of interest under a CMV promoter. None of the opsins investigated caused changes in retinal thickness; induced apoptosis in the retina or in transgene expressing cells; or reduced expression of PKCα (a specific bipolar cell marker). No increase in retinal inflammation at the level of gene expression (IBA1/AIF1) was found within the treated mice compared to controls. The expression of hRHO, CoChR or ReaChR under a strong constitutive promoter in retinal ON bipolar cells following intravitreal delivery via AAV2 does not cause either gross changes in retinal health, or have a measurable impact on the survival of targeted cells.

2020 ◽  
Vol 20 ◽  
Author(s):  
L. Hajba ◽  
A. Guttman

: Adeno-associated virus (AAV) is one of the most promising viral gene delivery vectors with long-term gene expression and disease correction featuring high efficiency and excellent safety in human clinical trials. During the production of AAV vectors,there are several quality control (QC)parameters that should be rigorously monitored to comply with clini-cal safety and efficacy. This review gives a short summary of the most frequently used AVV production and purification methods,focusing on the analytical techniques applied to determine the full/empty capsid ratio and the integrity of the encapsidated therapeutic DNA of the products.


2005 ◽  
Vol 52 (2) ◽  
pp. 293-299 ◽  
Author(s):  
Matthias H Kapturczak ◽  
Sifeng Chen ◽  
Anupam Agarwal

Relatively successful elsewhere, gene delivery aimed at the vasculature and kidney has made very little progress. In the kidney, the hurdles are related to the unique structure-function relationships of this organ and in the blood vessels to a variety of, mostly endothelial, factors making the delivery of transgenes very difficult. Among gene-therapeutic approaches, most viral gene delivery systems utilized to date have shown significant practical and safety-related limitations due to the level and duration of recombinant transgene expression as well as their induction of a significant host immune response to vector proteins. Recombinant adeno-associated virus (rAAV) vectors appear to offer a vehicle for safe, long-term transgene expression. rAAV-based vectors are characterized by a relative non-immunogenicity and the absence of viral coding sequences. Furthermore, they allow for establishment of long-term latency without deleterious effects on the host cell. This brief review addresses problems related to transgene-delivery to kidney and vasculature with particular attention given to rAAV vectors. The potential for gene therapy as a strategy for selected renal and vascular diseases is also discussed.


2002 ◽  
Vol 73 (2) ◽  
pp. 432-437 ◽  
Author(s):  
Uz Stammberger ◽  
Aima N. Uduehi ◽  
Bartosz Kubisa ◽  
Thierry Roth ◽  
Ralph A. Schmid

Biomaterials ◽  
2013 ◽  
Vol 34 (37) ◽  
pp. 9657-9665 ◽  
Author(s):  
Michael Keeney ◽  
Sheila Onyiah ◽  
Zhe Zhang ◽  
Xinming Tong ◽  
Li-Hsin Han ◽  
...  

Biomaterials ◽  
2011 ◽  
Vol 32 (23) ◽  
pp. 5402-5410 ◽  
Author(s):  
Stephany Y. Tzeng ◽  
Hugo Guerrero-Cázares ◽  
Elliott E. Martinez ◽  
Joel C. Sunshine ◽  
Alfredo Quiñones-Hinojosa ◽  
...  

2013 ◽  
Vol 14 (1) ◽  
pp. 46-60 ◽  
Author(s):  
Weiwei Wang ◽  
Wenzhong Li ◽  
Nan Ma ◽  
Gustav Steinhoff

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