# In situ evaluation of combinatorial gene regulation in the human genome

> **NIH NIH R01** · UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH · 2021 · $654,613

## Abstract

PROJECT SUMMARY
Gene expression enhancers are critical for establishing transcriptional programs in numerous contexts
throughout metazoans. There have been incredible advancements over the last decade in our ability to predict
enhancers and recent consortium efforts have identified millions of candidate enhancers across the human
genome. Unfortunately, it is difficult to test candidate enhancers at their endogenous locus with current
approaches requiring a large amount of effort to assess the activity of a single enhancer. Enhancers often
work in combination to regulate a single gene and current methods are not amenable to studying combinatorial
enhancer usage on a large-scale. To address the need for high-throughput methods for studying enhancers in
their native genomic context, we have adapted the CRISPR/Cas9 system to target enhancers and plan to test
the necessity and sufficiency of hundreds of genomic loci in driving transcriptional responses. By targeting
multiple distal regulatory regions simultaneously, we also have the ability to assess how enhancers work in
concert. During this project we will develop methods for high-throughput testing of enhancers in situ, using
estrogen receptor α (ER) and estrogen signaling in endometrial cancer cells as a model system. There are
three aims of the proposed project: (1) Determination of combinations of candidate enhancers that are
necessary for proper transcription regulation; (2) Identification of combinations of regulatory regions that are
sufficient to recapitulate an ER-driven transcriptional response; (3) Comparison of genetic deletion of
enhancers to epigenetic engineering. The successful completion of this project will lead to the discovery of
regulatory regions that are important for transcription regulation and provide an understanding of how
combinations of enhancers work in concert to regulate transcription. This project will also establish methods
for high-throughput testing of enhancers in their endogenous locus, which will be useful for studying enhancers
in a number of additional biological systems.

## Key facts

- **NIH application ID:** 10147743
- **Project number:** 5R01HG008974-05
- **Recipient organization:** UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH
- **Principal Investigator:** Jason Gertz
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $654,613
- **Award type:** 5
- **Project period:** 2017-07-01 → 2023-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10147743

## Citation

> US National Institutes of Health, RePORTER application 10147743, In situ evaluation of combinatorial gene regulation in the human genome (5R01HG008974-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10147743. Licensed CC0.

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