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Updated: Apr 23, 2026
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
Genome-wide CRISPR knockout screening identifies novel disease-associated genes in retinal pigment epithelium cells
1Genetic Diseases Key Laboratory of Sichuan Province, Department of Medical Genetics, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, China.
Abstract:
Dysfunction and degeneration of retinal pigment epithelium (RPE) cells are common pathological features observed in various retinal degenerative diseases. It has been proposed to treat these diseases by either protecting RPE cells or replacing them with new RPE cells derived from stem cells. However, the development of effective therapeutic strategies is still limited due to the insufficient understanding of the pathogenic factors involved in retinal degeneration and their impact on the function and survival of RPE cells. In this study, we employed genome-scale CRISPR knockout (KO) screening in human RPE cells to identify genes critical for RPE cell survival. Over 300 genes were identified, including well-established housekeeping genes as well as several candidate genes previously linked to retinal degeneration, many of which still lack comprehensive investigation. Among these, we further validated PRPF38B, which was both enriched in our screening and highlighted in a prior family-based linkage study, as essential for RPE cell survival, thus confirming the effectiveness of our approach. As a component of the spliceosome, we found that PRPF38B is crucial for functions specific to RPE cells, offering new insights into its role in retinal degeneration. Our study presents a novel approach for investigating risk genes associated with retinal diseases and may inspire future research on RPE cells and vision disorders.
Insights
Researchers identified over 300 genes crucial for retinal pigment epithelium (RPE) cell survival using CRISPR screening. PRPF38B was validated as essential for RPE survival, offering new insights into retinal degeneration.
Area of Science:
- Ophthalmology
- Genetics
- Cell Biology
Background:
- Retinal pigment epithelium (RPE) cell dysfunction is central to many retinal degenerative diseases.
- Current therapeutic strategies are limited by incomplete understanding of RPE cell degeneration factors.
- Identifying genes impacting RPE cell survival is crucial for developing new treatments.
Purpose of the Study:
- To identify genes essential for human RPE cell survival using genome-scale CRISPR knockout screening.
- To uncover novel pathogenic factors contributing to retinal degeneration.
- To validate candidate genes, such as PRPF38B, for their role in RPE cell survival.
Main Methods:
- Genome-scale CRISPR knockout (KO) screening was performed on human RPE cells.
- Over 300 genes critical for RPE cell survival were identified.
- PRPF38B was selected for further validation based on screening enrichment and prior linkage studies.
Main Results:
- The study identified over 300 genes, including housekeeping genes and potential retinal degeneration candidates.
- PRPF38B was confirmed as essential for RPE cell survival.
- PRPF38B's role as a spliceosome component crucial for RPE-specific functions was elucidated.
Conclusions:
- CRISPR screening is an effective approach for identifying genes involved in RPE cell survival and retinal diseases.
- PRPF38B is a key gene for RPE cell survival and a potential target for retinal degeneration therapies.
- This research provides new insights into the genetic basis of retinal degeneration and RPE cell function.
