# A combinatorial synthetic strategy for developing genome-editing protein-delivery agents targeting mouse retina

## TL;DR

Researchers developed a new delivery system using Coomassie lipidoids to efficiently deliver gene-editing proteins to the mouse retina, offering potential for treating inherited retinal diseases.

## Contribution

A novel non-viral delivery system using Coomassie brilliant blue-derived lipidoids for efficient and precise genome editing in the retina.

## Key findings

- Subretinal injection of Cre with CBB-lipidoids leads to robust recombination in retinal cells.
- CBB11 liposome-RNP complexes result in a 120-fold increase in base editing compared to RNP alone.
- Delivery of ABE RNP using CBB11 restores the scotopic ERG b-wave response in the rd12 mouse model.

## Abstract

CRISPR/Cas9-based gene-editing technologies offer promise for treating inherited retinal diseases (IRDs), however safe and efficient ocular delivery of precision editors remains challenging. To address this challenge, we report a class of Coomassie brilliant blue (CBB)-derived lipidoids that bind and deliver proteins. Subretinal injection of Cre complexed with these lipidoids into mT/mG mice leads to robust recombination in the retinal pigment epithelium and photoreceptors. We employ the CBB-lipidoid platform to deliver adenine base editor (ABE) ribonucleoproteins (RNP). Incorporating CBB lipidoids into liposomes improves delivery efficiency. CBB11 stands out for facilitating precise in vivo ABE-mediated gene editing. Delivery of liposome-CBB11-RNP complexes results in a 120-fold increase in base editing compared to RNP alone and restores the scotopic ERG b-wave response in the rd12 mouse model. These results demonstrate the potential of CBB-augmented, liposome-RNP systems for therapeutic gene editing in the eye, paving the way for single-dose precision medicines to treat IRDs.

Precise and efficient CRISPR genome editing requires specialized delivery systems. Here, the authors develop Coomassie lipidoids that deliver purified adenine base editors into retinal tissues, making it possible to achieve robust genome editing with a defined, non-viral nanomedicine.