# Determining the specificity and biological functions of widespread host mRNA degradation by RNase L

> **NIH NIH F32** · UNIVERSITY OF COLORADO · 2021 · $70,458

## Abstract

Project Summary/Abstract
The innate immune response is crucial for controlling infection by human pathogens. However, over-activation
of the innate immune response can cause chronic inflammation that leads to human diseases, such as cancers
and autoimmune disorders. To better understand and treat these diseases, developing a deeper understanding
of how the innate immune system functions is paramount. In particular, the mechanisms that lead to global host
shut-off of translation in response to double-stranded RNA (dsRNA), while allowing the expression of dsRNA-
induced antiviral and pro-inflammatory mRNAs has remained an incompletely understood aspect of the innate
immune response.
Assessment of the potent antiviral endoribonuclease, ribonuclease L (RNase L), at the single-cell level revealed
that it is the primary driver of translational arrest and functions by promoting rapid and widespread turnover of
mRNAs. This is a significant shift in the understanding of dsRNA-induced translational arrest, as it would permit
translation of mRNAs that are not degraded by RNase L. Consistent with this, the mRNA of the potent antiviral
interferon-b (IFN-b) cytokine escapes RNase L-mediated mRNA turnover, potentially allowing for translation of
the IFN-b mRNA.
Based on these preliminary findings, this application proposes to test the hypothesis that widespread RNase L-
mediated mRNA turnover functions to preferentially promote translation of antiviral mRNAs that are resistant to
RNase L-mediated mRNA turnover. These findings may provide novel insights into RNase L-mediated
translational arrest and antiviral gene expression that will have translational importance for understanding and
treating human disease associated with dysregulation of the innate immune response. Aim 1: High-throughput
sequencing and single-molecule fluorescent in situ hybridization (smFISH) will be used to identify mRNAs in
addition to the IFN-b mRNA that are resistant to RNase L-mediated mRNA turnover. Aim 2: Targeted
mutagenesis, chimeric mRNAs, and heterologous promoters, will be used to determine the mechanistic basis by
which RNase L resistant mRNA escape RNase L-mediated mRNA turnover. Aim 3: Single-cell analysis of mRNA
expression and protein translation in conjunction with ribosomal profiling will be performed to determine if RNase
L-mediated mRNA promotes the translation of RNase L resistant mRNAs. Completion of these aims will
determine the breadth of mRNAs resistant to RNase L-driven mRNA turnover, determine the mechanism(s) by
which mRNAs escape RNase L-mediated mRNA turnover, and provide a novel mechanism by which RNase L
regulates antiviral gene expression during the innate immune response.
!

## Key facts

- **NIH application ID:** 10116269
- **Project number:** 5F32AI145112-03
- **Recipient organization:** UNIVERSITY OF COLORADO
- **Principal Investigator:** James M Burke
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $70,458
- **Award type:** 5
- **Project period:** 2019-03-09 → 2022-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10116269

## Citation

> US National Institutes of Health, RePORTER application 10116269, Determining the specificity and biological functions of widespread host mRNA degradation by RNase L (5F32AI145112-03). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10116269. Licensed CC0.

---

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