# A Universal DNA Hemimethylation Probe

> **NIH NIH R21** · CARNEGIE INSTITUTION OF WASHINGTON, D.C. · 2020 · $444,208

## Abstract

PROJECT SUMMARY
Cytosine DNA methylation is a post-replicative epigenetic modification that regulates the organization and
expression of the genetic information. Deficiencies in DNA methylation associate with numerous diseases,
most notably genomic imprinting disorders, immunodeficiencies, and cancers. In the vertebrates, cytosine
methylation happens in the CpG context. DNA replication produces a short-lived hemimethylated DNA
intermediate. Due to its fleeting nature and the lack of adequate tools, DNA hemimethylation remains
underexplored. Intriguingly, recent studies revealed fascinating examples of persistent DNA hemimethylation.
These studies suggest a broader spectrum of functional states of DNA methylation. To begin to address this
gap in our understanding of this potentially crucial element of the epigenetic regulation of the genome, we
propose to adapt a SET- and RING Finger-Associated (SRA) domain of UHRF1 to generate a DNA
hemimethylation probe for in vitro and in vivo studies. The compact SRA domain is self-sufficient to bind
hemimethylated DNA with high specificity. To accomplish the overarching goal of this proposal, we will first
produce recombinant SRA domain proteins and develop methods for the isolation and visualization of
hemimethylated DNA in complex genomes in vitro. To study DNA hemimethylation in vivo, we will express a
fusion protein comprised of the SRA domain and mScarlet fluorescent protein in mouse embryos, embryonic
stem cells, and transgenic mice. We will use live imagining of cells with hemimethylated DNA to validate
chromatin localization patterns and dynamics of the SRA-mScarlet fusion protein. This research will enhance
our understanding of the epigenetic regulation of the genome in development and disease.

## Key facts

- **NIH application ID:** 10038450
- **Project number:** 1R21HG011282-01
- **Recipient organization:** CARNEGIE INSTITUTION OF WASHINGTON, D.C.
- **Principal Investigator:** ALEX BORTVIN
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $444,208
- **Award type:** 1
- **Project period:** 2020-08-01 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10038450

## Citation

> US National Institutes of Health, RePORTER application 10038450, A Universal DNA Hemimethylation Probe (1R21HG011282-01). Retrieved via AI Analytics 2026-05-31 from https://api.ai-analytics.org/grant/nih/10038450. Licensed CC0.

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