Targeted Transgene Expression in Müller Glia of Normal and Diseased Retinas Using Lentiviral Vectors

Abstract

Pseudotyped LV vectors containing glia-specific promoters efficiently transduce and direct sustained transgene expression in retinal Müller glia. Vectors of this type will be useful for experimental treatment of retinal disease, as well as for physiological and developmental investigations of the retina.

AAV Mediated GDNF Secretion From Retinal Glia Slows Down Retinal Degeneration in a Rat Model of Retinitis Pigmentosa

“…Application of the AAV variant ShH10, which was previously created via the directed evolution of AAV to better infect glial cells, 13 results in selective targeting of glia and minimizes ectopic expression in neurons (see Supplementary Figure S4), which contrasts to the tropism of most natural AAV serotypes that preferentially transduce neurons with minimal glial transduction. [22][23][24] In most previous work, restricting gene expression to a given cell type was achieved primarily via incorporation of cell-specific promoters, 25,26 leading in most cases to weaker expression than can be achieved with strong, ubiquitous promoters. In contrast, transductional targeting at the initial step of viral interaction with receptors on the cell surface minimizes the loss of vector genomes to uptake by off-target cells that do not express the transgene.…”

Let There Be Light: Gene and Cell Therapy for Blindness

“…It is the only vector that can provide gene delivery to the inner retina after intravitreal delivery. [20][21][22] Although the small size of the AAV particle is an advantage, it is also its weakness: the 25-nm AAV particles can package only 4.7 kb of genetic material, limiting its application in some diseases. Nevertheless, AAV has yet another advantage that has contributed to its development as a gene delivery vehicle: it is a nonenveloped virus and its capsid can be easily modified using genetic engineering techniques.…”

Intravitreal inhibition of complement C5a reduces choroidal neovascularization in mice

“…Volumes of 2 μl were applied intravitreally using a 33-gauge needle (World Precision Instruments, Sarasota, FL, USA) attached to a 10-μl glass syringe (Hamilton Bonaduz, Bonaduz, Switzerland), which is an established method [21,22]. To investigate the influence of different concentrations, four groups consisting of ten animals each were analyzed.…”