# Identification of gene regulatory networks that control proliferative and neurogenic competence in mammalian Müller glia

> **NIH NIH R01** · JOHNS HOPKINS UNIVERSITY · 2020 · $505,313

## Abstract

Project Summary
Müller glia of cold-blooded vertebrates can re-enter the cell cycle and rise to photoreceptors following retinal
injury, while mammals have lost this ability. As part of the NEI Audacious Goals Initiative, we have conducted a
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comprehensive analysis of injury-induced changes in gene expression and chromatin accessibility in zebrafish,
chick and mouse M ller glia, allowing us to identify both evolutionarily consüerved and species-specific gene
regulatory networks that regulate glial reprogramming. This has identified a set of dedicated gene regulatory
networks in mice tühat restrict proliferative and neurogenic competence in M ller glia. We aim to use these
findings to gain a more complete insight into the molecular mechanisms that regulate neurogenic competence
in mammalian M ller glia, and to develop treatments that can maximize generation of glial-derived
photoreceptors while simultaneously not depleting the number of existing glia. To do this, we propose to
generate individual loss of function mutants of the top candidate negative regulators of proliferative and
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neurogenic competence using AAV-mediated CRISPR/Cas9 gene disruption. We will first validate efficacy of
sgRNAs targeting individual TFs, comprehensively profile chanüges in geüne expression in reactive M ller glia
following loss of function of these genes, and characterize the fate of M ller glia-derived cells. We will then
conduct combinatorial loss of function of negative regulators of M ller glia reprogramming to enhance generation
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of glial-derived retinal progenitor cells in wildtype and Nfia/b/x-deficient mice. Finally, we will combine CRISPR-
mediated loss of function analysis with overexpression of Ascl1, Crx and Nrl to enhance generation of M ller
glia-derived rod photoreceptors in both wildtype and dystrophic retina. We predict that these studies may
substantially advance cell-based regenerative treatments aimed at restoring retinal photoreceptors lost due to
blinding diseases.

## Key facts

- **NIH application ID:** 10029171
- **Project number:** 1R01EY031685-01
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Seth Blackshaw
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $505,313
- **Award type:** 1
- **Project period:** 2020-08-01 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10029171

## Citation

> US National Institutes of Health, RePORTER application 10029171, Identification of gene regulatory networks that control proliferative and neurogenic competence in mammalian Müller glia (1R01EY031685-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10029171. Licensed CC0.

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