# CRISPR/Cas9 approaches to model exfoliation syndrome

> **NIH NIH R21** · UNIVERSITY OF IOWA · 2020 · $193,125

## Abstract

Exfoliation syndrome is common and may affect 5-10% of Americans and up to 50% in some populations (i.e.
Icelanders). As many as 10 to 50% of those with exfoliation syndrome develop high intraocular pressure and a
secondary glaucoma - exfoliative glaucoma, which causes blindness and visual disability in millions worldwide.
Exfoliative syndrome has a strong genetic basis, but complex inheritance. Genome-wide association studies
(GWAS) of exfoliation syndrome have successfully identified 7 genetic factors that increase risk for disease:
LOXL1, CACNA1A, POMP, TMEM136, AGPAT1, RBMS3, and SEMA6A. However, an animal model has not
yet been built based upon these gene discoveries. There is a critical need for these human GWAS discoveries
to be translated into animal models.
We previously reported mice, B6-Lystbg-j, with a mutation in the Lyst gene, that have some features of
exfoliation syndrome, but produce only tiny amounts of exfoliation material and do not develop glaucoma.
We hypothesize that inactivating the 7 exfoliation syndrome risk factor genes using CRISPR/Cas9
methods will generate animal models of exfoliation syndrome and glaucoma. We further hypothesize that
introducing these mutations on the B6-Lystbg-j background strain will take advantage of its subtle exfoliation
syndrome features and facilitate developing a new line with pronounced features, high pressure and glaucoma.
We have created CRISPR reagents to inactivate the seven exfoliation syndrome risk factors and we have
begun injecting mouse embryos. We are injecting pools of CRISPR reagents for our target genes in order to
get founder mice with one or more inactivated risk factor genes. All mutations are made in B6-Lystbg-j mice.
AIM 1A: ASSESS FOUNDER MICE. We will assess the eyes of founder mice (in vivo) that have one or more
inactivated exfoliation syndrome risk factor genes for exfoliation material. Those with exfoliative material in
their eyes will be aged to 1 year, assessed for high intraocular pressure, and glaucomatous optic neuropathy.
AIM 1B: GENERATE MICE WITH MULTIPLE INACTIVATED RISK FACTORS. We will conduct crosses to
produce mice with multiple inactivated exfoliation syndrome risk factor genes on a B6-Lystbg-j background:
1) To produce mice with 2 heterozygously inactivated risk factor genes (with extreme individual phenotypes).
2) To produce mice with 2 risk factors (Loxl1, Cacn1a1) heterozygously inactivated (top GWAS hits).
AIM 2A DETERMINE EXPRESSION PATTERNS OF RISK FACTOR GENES IN B6-Lystbg-j MICE. We will
assess expression of the 7 risk factor genes at the mRNA and protein level in ocular tissues of B6-Lystbg-j mice
relevant to exfoliation syndrome pathophysiology (lens, iris, and iridocorneal angle).
AIM 2B DETERMINE EXPRESSION PATTERNS OF RISK FACTOR GENES IN OUR CRISPR/Cas9 MICE.
INNOVATION. Pooling CRISPR guide RNAs to make mice with inactivated risk factor genes (alone and in
combination). Creating the 1st animal model of exfoliation syndrome from GWAS hits...

## Key facts

- **NIH application ID:** 9852602
- **Project number:** 5R21EY029991-02
- **Recipient organization:** UNIVERSITY OF IOWA
- **Principal Investigator:** Michael G Anderson
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $193,125
- **Award type:** 5
- **Project period:** 2019-02-01 → 2021-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9852602

## Citation

> US National Institutes of Health, RePORTER application 9852602, CRISPR/Cas9 approaches to model exfoliation syndrome (5R21EY029991-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9852602. Licensed CC0.

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