# Developing single nuclei tRNA-seq to profile tRNA diversity at the single cell level

> **NIH NIH R21** · CASE WESTERN RESERVE UNIVERSITY · 2024 · $442,750

## Abstract

Project Summary/Abstract
Transfer RNA (tRNA) biogenesis is essential for determining the pool of tRNAs that set the translational state of
cells, and defects in the tRNA biogenesis machinery are associated with several types of neurodegenerative
diseases. While bulk tRNA-sequencing assays of tissues and cell cultures have revealed that tRNA expression
and modification are regulated in tissue and disease-specific manners, technology to date has hindered our
ability to define tRNA processing, expression, or modification changes in neurodevelopment and
neurodegeneration at the level of single cells, limiting important insight and therapeutic development. In this
application, we propose to develop and apply a novel tRNA sequencing method to stem cell derived
neurons and cerebral organoids to reveal complex tRNA processing and modification patterns at the
single cell level for the first time. We will utilize preexisting laboratory models to establish and optimize our
technology (Aim 1) before profiling cerebral organoids with pathogenic variants in CLP1 and isogenic controls
(Aim 2). These findings will reveal the complexity of tRNA processing and modification at unprecedented cellular
level resolution. In addition to neurodevelopment and neurodegeneration, our research will have broad impacts
on the study of tRNAs across all tissues, as it will be applicable to any frozen or fresh tissue source. Outside of
neurobiology, it will also benefit other fields such as cancer biology and virology where tRNA modulation is known
to be important.

## Key facts

- **NIH application ID:** 10995456
- **Project number:** 1R21NS139431-01
- **Recipient organization:** CASE WESTERN RESERVE UNIVERSITY
- **Principal Investigator:** Ashleigh E Schaffer
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $442,750
- **Award type:** 1
- **Project period:** 2024-07-15 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10995456

## Citation

> US National Institutes of Health, RePORTER application 10995456, Developing single nuclei tRNA-seq to profile tRNA diversity at the single cell level (1R21NS139431-01). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10995456. Licensed CC0.

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