# Elucidating the cis-regulatory grammar of human photoreceptors

> **NIH NIH R01** · WASHINGTON UNIVERSITY · 2021 · $528,530

## Abstract

Project Summary
 One of the major unsolved challenges of the genomic era is to understand how individual sequence
variation contributes to disease. Coding variation is increasingly well understood, but our knowledge of the
effects of non-coding variation remains rudimentary. The goal of this proposal is to develop a platform for the
analysis of non-coding cis-regulatory variation in human retinal disease. We hypothesize that sequence
variants in photoreceptor-specific cis-regulatory elements (CREs; e.g., enhancer/promoters) play an important
role in retinal disease by altering the expression levels of disease-related genes. Currently, assessing the
effects of variants that fall within non-coding DNA represents a challenging problem, in part, because our ability
to assay CREs in a high-throughput fashion is limited. To address this challenge, we have developed a
technique called CRE-seq (Cis-Regulatory Element analysis by sequencing). In CRE-seq, individual CREs are
fused to reporter genes, each containing a unique DNA barcode. The resultant CRE-reporter library, consisting
of thousands of constructs, is introduced into living retina, and reporter gene expression is quantified by
counting barcoded transcripts with RNA-seq. CRE-seq promises to revolutionize our ability to measure the
effects of human cis-regulatory variants. To achieve this goal, we propose three Specific Aims. In Aim 1, we
will use ATAC-seq to identify candidate CREs in both developing and mature human photoreceptors. In Aim 2,
we will utilize a combination of computational and experimental approaches (including CRE-seq analysis) to
analyze the CREs identified in Aim 1 and thereby elucidate the cis-regulatory grammar of human
photoreceptors. CRE-seq will be performed in both mouse retinas as well as ES cell-derived human retinal
organoids. These studies will provide the first comprehensive view of the human photoreceptor 'cis-regulome'
and will begin to decipher the cis-regulatory code of human photoreceptors. In Aim 3, we will use a
‘mutagenesis and screening’ approach to engineer a library of compact (150 bp), highly active enhancers for
targeting human photoreceptors in gene therapy applications. If successful, these studies will establish a
quantitative platform for the analysis of non-coding cis-regulatory variation, thereby enabling comprehensive
interpretation of whole-genome sequence data in the context of retinal disease. In addition, they will engineer a
suite of new enhancers for human retinal gene therapy.

## Key facts

- **NIH application ID:** 10135089
- **Project number:** 5R01EY030075-02
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** JOSEPH CORBO
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $528,530
- **Award type:** 5
- **Project period:** 2020-04-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10135089

## Citation

> US National Institutes of Health, RePORTER application 10135089, Elucidating the cis-regulatory grammar of human photoreceptors (5R01EY030075-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10135089. Licensed CC0.

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