# Investigating a novel role of ejaculate RNA in fertility

> **NIH NIH R21** · UNIVERSITY OF ARIZONA · 2020 · $183,958

## Abstract

Project Summary/Abstract
 The male ejaculate of internally fertilizing organisms is a complex mixture of diverse components including
sperm and numerous other macromolecules. Research on male contributions to fertility has mainly focused on sperm, and,
more recently on proteins present in the seminal fluid. Much less attention has been directed toward understanding the
role that other ejaculate molecules might play in fertilization success. The goal of the proposed research is to investigate
the functional significance of ejaculate RNA, which is found in the ejaculates of diverse organisms ranging from humans
to insects, though little is known about its potential function. Specifically, the proposed research involves three specific
aims, which together evaluate whether coding and long non-coding RNA (lncRNA) in the ejaculate of male Drosophila
arizonae has functional effects on fertility either by directing the production of key proteins or through regulatory effects
within the female reproductive tract. Aim one combines ribosome profiling (ribo-seq) and proteomics to assess whether
male mRNAs are translated within females. The challenge of differentiating male and female RNA and proteins within
the female reproductive tract is overcome by exploiting natural genetic variation in the D. arizonae study system and by
the adoption of recently developed methods for metabolically labeling the proteome of adult Drosophila using stable
isotope labeled amino acids. Because lncRNAs often function through interactions with protein, aim two involves the
identification of RNA-protein interactions using RNA Antisense Purification with Mass Spectrometry (RAP-MS).
Interacting proteins with annotated functions will provide key insight into the mechanisms by which lncRNAs function.
Aim three proposes to generate knockout male lines for a select number of RNAs using the CRISPR genome editing
system. Reproductive output of crosses between knockout males and wild-type females will be compared to wild-type
crosses, with the expectation that if a male RNA is involved in fertility, crosses involving knockout males will have lower
reproductive output than crosses with wild-type males. Altogether the results of the proposed studies will provide novel
insights into the molecular interactions between male and female molecules that ultimately determine fertilization success.
Moreover, this research may have important implications for identifying the underlying causes of idiopathic infertility, the
development of assistive reproductive techniques, and the design of new strategies for integrative pest management.

## Key facts

- **NIH application ID:** 9873987
- **Project number:** 5R21HD097545-02
- **Recipient organization:** UNIVERSITY OF ARIZONA
- **Principal Investigator:** Jeremy Matthew Bono
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $183,958
- **Award type:** 5
- **Project period:** 2019-02-15 → 2023-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9873987

## Citation

> US National Institutes of Health, RePORTER application 9873987, Investigating a novel role of ejaculate RNA in fertility (5R21HD097545-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9873987. Licensed CC0.

---

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