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Related Experiment Video

Updated: Jun 15, 2025

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

Published on: April 19, 2015

Retinal Penetrating Adeno-Associated Virus.

Binit Kumar¹, Manish Mishra¹, Siobhan Cashman¹

¹Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, Massachusetts, United States.

Investigative Ophthalmology & Visual Science

|August 22, 2024

Summary

A novel adeno-associated virus (AAV) vector incorporating a truncated molecular chaperone (IKV) successfully delivered genes to the outer retina, including photoreceptors and RPE, via intravitreal injection in mice.

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Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

Published on: August 7, 2015

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Last Updated: Jun 15, 2025

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

Published on: April 19, 2015

Subconjunctival Administration of Adeno-associated Virus Vectors in Small Animal Models

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Published on: March 16, 2022

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

Published on: August 7, 2015

Area of Science:

Ophthalmology
Gene Therapy
Molecular Biology

Background:

Current gene delivery to the outer retina commonly uses subretinal injection of adeno-associated virus (AAV).
Intravitreal injections are a more common and less invasive method for retinal drug delivery.
Developing intravitreal AAV vectors is crucial for advancing retinal gene therapy.

Purpose of the Study:

To engineer AAV vectors capable of reaching the outer retina following intravitreal administration.
To assess the efficacy of modified AAV vectors for gene delivery to retinal cells.

Main Methods:

A molecular chaperone (Nuc1) and its truncated version (IKV) were genetically incorporated into AAV2/9 VP1 protein.
Recombinant AAV vectors encoding GFP or Nrf2 were constructed with IKV modification.
These vectors were administered via intravitreal injection into mice, and gene expression and oxidative stress markers were quantified.

Main Results:

AAV2/9 vectors with full Nuc1 did not yield significant retinal gene expression.
AAV2/9 vectors with the IKV modification demonstrated robust green fluorescent protein (GFP) expression in photoreceptors and retinal pigment epithelium (RPE) after intravitreal injection.
Intravitreal delivery of Nrf2 using IKV-modified AAV vectors reduced oxidative stress in the retina.

Conclusions:

A novel AAV vector incorporating IKV enables efficient gene delivery to the outer retina (photoreceptors and RPE) via intravitreal injection in a murine model.
This engineered vector represents a promising advancement for non-invasive gene therapy strategies targeting retinal diseases.