# Reactivation of Inactivated X-linked Genes Via Inhibition of Histone Demethylase KDM5C

> **NIH NIH F31** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2020 · $38,131

## Abstract

Project Summary/Abstract
X-chromosome inactivation equalizes X-linked gene expression between XX female and XY male mammals. X-
inactivation is separated into two distinct phases: an initiation phase and a maintenance phase. The Kalantry lab
has recently found that the X-linked gene, Kdm5c, is both necessary and sufficient to cause initiation of X-
inactivation in a dose-dependent manner. KDM5C is a demethylase enzyme that removes the histone
H3K4me2/3 chromatin marks which are associated with active transcription. Deleting both copies of Kdm5c
abrogates the initiation of X-inactivation. Deleting one copy of Kdm5c leads to deficient silencing of a subset of
X-linked genes when X-inactivation initiates. The specific goal of my project is to determine the dose-
dependent role of KDM5C in maintaining X-inactivation. In Aim 1 of this proposal, I will test a role for KDM5C
in maintaining X-inactivation. I will generate hybrid primary cortical cells, which are in the maintenance phase of
X-inactivation, with biased X-inactivation from mouse embryos with a conditional Kdm5c mutation on the active-
X. By deleting one Kdm5c allele in the cortical neurons, I will test which X-linked genes require KDM5C to remain
silenced through both high- and low-throughput approaches. In Aim 2 of this proposal, I will inhibit KDM5C protein
by administering a validated KDM5 pharmacological inhibitor to cultured hybrid primary cortical neurons. I will
test KDM5C inhibition by chromatin immunoprecipitation (ChIP) for H3K4me2/3 followed by next-generation
sequencing (Seq). I will also test X-linked gene expression in the KDM5C-inhibited cortical neurons by allele-
specific RNA-Seq. Through these experiments, I will learn how X-inactivation occurs and is maintained. The
expected findings also promise a potential therapy for females with X-linked disorders.

## Key facts

- **NIH application ID:** 9989388
- **Project number:** 1F31HD102122-01
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Megan Brianne Trotter
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $38,131
- **Award type:** 1
- **Project period:** 2020-09-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9989388

## Citation

> US National Institutes of Health, RePORTER application 9989388, Reactivation of Inactivated X-linked Genes Via Inhibition of Histone Demethylase KDM5C (1F31HD102122-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9989388. Licensed CC0.

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