New publication: Study of genes contributing to age-related macular degeneration

A/Prof Wong's team has used CRISPRi technology to study the function of gene contributing to AMD patholobiology.

Development of a CRISPRi Human Retinal Pigmented Epithelium Model for Functional Study of Age-Related Macular Degeneration Genes

Jiang-Hui Wang, Daniel Urrutia-Cabrera, Jarmon G Lees, Santiago Mesa Mora, Tu Nguyen, Sandy SC Hung, Alex W Hewitt, Shiang Y Lim, Thomas L Edwards, Raymond CB Wong

International Journal of Molecular Sciences, 2023, accepted 3 Feb 2023, (IF = 6.21)

Abstract

Age-related macular degeneration (AMD) is a blinding disease characterised by dysfunction of the retinal pigmented epithelium (RPE) which culminates in disruption or loss of the neurosensory retina. Genome-wide association studies have identified >60 genetic risk factors for AMD; however, the expression profile and functional role of many of these genes remain elusive in human RPE. To facilitate functional studies of AMD-associated genes, we developed a human RPE model with integrated CRISPR interference (CRISPRi) for gene repression by generating a stable ARPE19 cell line expressing dCas9-KRAB. We performed transcriptomic analysis of the human retina to prioritise AMD-associated genes and selected TMEM97 as a candidate gene for knockdown study. Using specific sgRNAs, we showed that knockdown of TMEM97 in ARPE19 reduced reactive oxygen species (ROS) levels and exerted a protective effect against oxidative stress-induced cell death. This work provides the first functional study of TMEM97 in RPE and supports a potential role of TMEM97 in AMD pathobiology. Our study highlights the potential for using CRISPRi to study AMD genetics, and the CRISPRi RPE platform generated here provided a useful in vitro tool for functional studies of AMD-associated genes.

Read the publication here.