# Control of spermatogonial stem cell formation

> **NIH NIH R01** · UNIVERSITY OF MINNESOTA · 2020 · $23,475

## Abstract

Abstract / Project Summary
 The overall objective of the proposed work is to understand how the Dmrt1 gene controls
spermatogonial stem cell formation in the mammalian testis. The testis has two essential functions: production
of sperm, the cells that serve as vehicles for the immortality of male germ line DNA; and production of
hormones that direct other parts of the body to develop in a male-specific manner. Failures of these processes
cause infertility, germ cell cancer, and disorders of sex development (DSD). Dmrt1 belongs to a family of
conserved transcriptional regulators and it controls multiple crucial processes in the mammalian testis, both in
germ cells and somatic cells. A recent discovery is that DMRT1 is required to form spermatogonial stem cells
(SSCs). The central hypothesis of this proposal is that DMRT1 acts as a pioneer transcription factor to open
chromatin and allow other cooperating transcription factors to bind and regulate gene expression, thereby
orchestrating SSC formation. This proposal has three aims focused on a deeper understanding of how
DMRT1 controls SSC cell fate. Aim 1 asks how DMRT1 directs SSC formation. It examines how loss of Dmrt1
affects key events in SSC formation including proliferation and expression of SSC regulators. It then seeks a
mechanistic understanding of how DMRT1 regulates target gene transcription to control cell fate, testing the
hypothesis that DMRT1 is a pioneer transcription factor. The experiments will employ an array of state-of-the-
art genomic tools including ChIP-seq and ATAC-seq to find key regulatory targets and learn how DMRT1
binding in SSCs affects enhancer activity. Motif searches and ChIP will be used to identify likely cooperating
transcription factors. Aim 2 will identify the genes regulated directly and indirectly by DMRT1 during SSC
formation. Regulated transcripts will be identified by standard and single-cell RNA-seq and HiChIP will be
used to link regulatory regions to one another and to the transcriptional start sites they control. Aim 3 will test
the functional importance of selected DMRT1 cooperating transcription factors and target genes, using
lentiviral knockdown in cultured SSCs followed by in vivo validation for top candidates. The proposed work
has direct human health relevance: DMRT1 in humans is linked to infertility, testicular germ cell cancer and
DSD including male-to-female sex reversal. As a result, the proposed work will help uncover the mechanistic
basis of SSC formation and may provide general insights into stem cell biology.

## Key facts

- **NIH application ID:** 10130752
- **Project number:** 3R01GM127379-02S1
- **Recipient organization:** UNIVERSITY OF MINNESOTA
- **Principal Investigator:** David A. Zarkower
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $23,475
- **Award type:** 3
- **Project period:** 2019-01-01 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10130752

## Citation

> US National Institutes of Health, RePORTER application 10130752, Control of spermatogonial stem cell formation (3R01GM127379-02S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10130752. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*