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Prime Editing Strategy to Install the RPE65 c.1430A>G Dominant Mutation.

Bruna Lopes da Costa^1,2, Salvatore Marco Caruso^1,2, Yi-Ting Tsai^1,2

¹Department of Biomedical Engineering, Columbia University, New York, NY, USA.

Advances in Experimental Medicine and Biology

|February 10, 2025

Summary

This summary is machine-generated.

Researchers developed a prime editing strategy to model a rare form of inherited blindness caused by mutations in the retinal pigment epithelium 65-kDa protein (RPE65). This method enables the study of RPE65-mediated autosomal dominant retinitis pigmentosa using patient-derived cells.

Keywords:

Autosomal dominant retinitis pigmentosa (adRP)Prime editing RPE65

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Area of Science:

Ophthalmology
Genetics
Molecular Biology

Background:

The retinal pigment epithelium 65-kDa protein (RPE65) is crucial for the visual cycle.
Mutations in RPE65 typically cause autosomal recessive forms of inherited retinal diseases.
A patient with a rare autosomal dominant form of retinitis pigmentosa (adRP) linked to RPE65 was identified.

Purpose of the Study:

To develop a method for modeling RPE65-mediated adRP.
To establish a strategy for introducing specific RPE65 mutations into cells for disease modeling.
To create a platform for screening gene editing therapies for RPE65-adRP.

Main Methods:

Developed a prime editing strategy to precisely install the c.1430A>G mutation in RPE65.
Utilized prime editing in HEK293T cells and induced pluripotent stem cells (iPSCs).
Aimed to generate patient-specific iPSC-derived RPE cells for disease modeling.

Main Results:

Successfully developed and demonstrated a prime editing strategy for targeted RPE65 mutation installation.
The strategy allows for the precise introduction of the pathogenic c.1430A>G mutation.
This method is suitable for creating cellular models of RPE65-mediated adRP.

Conclusions:

Prime editing provides an effective strategy for modeling rare genetic mutations like those in RPE65.
This approach facilitates the study of RPE65-mediated adRP pathobiology.
The developed method supports the development and screening of gene editing therapies for inherited retinal diseases.