# Developing C. elegans as a model to understand tRNA-fragment biogenesis and function

> **NIH NIH R35** · CHILDREN'S HOSP OF PHILADELPHIA · 2024 · $445,000

## Abstract

Project Summary
Small RNAs are ubiquitous regulators of eukaryotic gene expression in nearly all aspects of physiology
from plants to humans. They function to regulate all facets of gene expression including transcription,
RNA stability, and translation. An emerging class of small regulatory RNAs are tRNA-fragments (tRFs),
produced from nucleolytic cleavage of tRNAs. tRFs have been implicated in cancer, neurodegenerative
disease, viral infection, fertility, epigenetic inheritance, and aging. While the biogenesis of these RNAs is
poorly understood, tRFs have been demonstrated to play roles in the regulation of transcription,
posttranscriptional regulation of mRNA stability, and translation. Yet, the functions of tRFs throughout the
different tissues of an organism in regulating cellular physiology are unknown.
 My lab is developing the roundworm C. elegans (worms) as a model to dissect the molecular
mechanisms underlying the biogenesis tRF and cellular functions throughout the animal. The robust
genetics, physiological assays, and molecular tools available in C. elegans provide a system ripe for the
discovery of tRF biology. Importantly, we have developed small RNA-sequencing techniques to detect
abundant tRFs in C. elegans, at levels much higher than previously published small RNA-seq datasets.
Using these techniques, we will characterize tRFs in the different tissues of the fully developed adult
using bulk and single-cell small RNA-seq. After determining the spatial and temporal expression of tRF
species we will utilize both forward and reverse genetics to determine new factors required for their
biogenesis. Further, we will employ biochemical enrichment strategies to determine factors that interact
with and affect tRF function. Upon determining genes required for tRF biogenesis and function we will
use mutant alleles of these factors to determine what roles these molecules have in regulating the
physiology of the organism. Finally, we will use worm strains carrying loss-of-function mutations in RNA
modifying enzymes to determine how RNA modifications affect tRF biogenesis, functions, and further
regulate the physiology of the animal. The work on tRFs in C. elegans in my lab represents the first
systematic dissections of all aspects of tRFs biology in a whole organism. What we learn about tRF
biology in worms will be used to generate hypotheses about tRFs in other organisms. As tRFs have been
implicated in many aspects of normal physiology in organisms ranging from yeast to humans, as well as
in a wide range of diseases, comprehensively understanding how tRF are regulated and function
represents a highly under-addressed aspect of biology.

## Key facts

- **NIH application ID:** 10875671
- **Project number:** 5R35GM151087-02
- **Recipient organization:** CHILDREN'S HOSP OF PHILADELPHIA
- **Principal Investigator:** Colin Conine
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $445,000
- **Award type:** 5
- **Project period:** 2023-07-01 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10875671

## Citation

> US National Institutes of Health, RePORTER application 10875671, Developing C. elegans as a model to understand tRNA-fragment biogenesis and function (5R35GM151087-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10875671. Licensed CC0.

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