# CRSIPR screening for novel regulators of retinal ganglion cell survival and axonal regeneration

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2020 · $533,682

## Abstract

Abstract/Project Summary
Neurodegenerative diseases including glaucoma are characterized by neuronal death and failure of damaged
axons to regenerate. Optic nerve crush (ONC), which transects all retinal ganglion cell (RGC) axons, is often
used to model this process, and to seek interventions that protect RGCs and promote regeneration. Following
ONC in mice, ~80% of the RGCs die within 2 weeks, and virtually none of the survivors regenerate axons.
We and others have used ONC to identify interventions that lead to increased survival, increased regeneration
or both. However, these treatments are only partially effective. For example, PTEN deletion increases RGC
survival by only two-fold, and of >45 RGC types, only a few (alpha-RGCs) extend axons. Additionally, the
growth rates of regenerating axons are slow and, most important, the regenerating axons rarely reach their
targets in the brain. It is therefore important to identify additional and improved promoters of survival and
regeneration. We will address this challenge by conducting an unbiased loss-of-function screen using
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats). Based on previous work from us and
others showing that knockdown-regulation of several transcription factors can improve survival, regeneration
or both, we have generated an AAV2-based sgRNA library for 1629 transcription factor genes; optimized
methods for delivering the sgRNAs and Cas9 to RGCs; and validated our ability to identify genes that regulate
survival and regeneration. In the proposed study, we will screen the entire library to find novel repressors of
programs required for neuroprotection and regeneration. For selected positive hits, we will identify RGC
subtypes that are protected and/or undergo axon regeneration after individual gene knockout. Finally, as a
first step to test identified candidates in a clinically relevant setting, we will choose three gRNAs with robust
neuroprotective effects and test their ability to protect RGCs in a widely-used glaucoma model. We expect
these studies will provide insights that will enable development of novel neuroprotective and regeneration-
promoting strategies for traumatic injury and glaucoma as well as other neurodegenerative diseases.

## Key facts

- **NIH application ID:** 9920148
- **Project number:** 5R01EY030204-02
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** ZHIGANG HE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $533,682
- **Award type:** 5
- **Project period:** 2019-05-01 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9920148

## Citation

> US National Institutes of Health, RePORTER application 9920148, CRSIPR screening for novel regulators of retinal ganglion cell survival and axonal regeneration (5R01EY030204-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9920148. Licensed CC0.

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