# Mouse models for live-cell imaging of endogenous Evf2 lncRNA

> **NIH NIH R03** · NORTHWESTERN UNIVERSITY · 2021 · $79,917

## Abstract

Abstract
 Long non-coding RNAs (lncRNAs) play critical roles in diverse cellular processes,
including transcriptional and post-transcriptional regulation. The Evf2 ultraconserved
lncRNA is a transcription regulating lncRNA transcribed from the Dlx5/6 ultraconserved
enhancer (UCE) in the developing forebrain. Unlike lncRNAs involved in imprinting, that
regulate gene expression in one direction, Evf2 positively and negatively regulates target
genes. Evf2 forms a ribonucleoprotein complex (Evf2-RNP) with 87 proteins and controls
the expression of a gene regulatory network that affects interneuron diversity and seizure
susceptibility, through mechanisms that involve the recruitment of transcription factors,
and the control of epigenetic modifications and chromosome topology.
 The study of the subnuclear localization of Evf2 is key for understanding its
function. Evf2 forms one or two RNA clouds per nucleus. A subset of Evf2-RNP proteins
localize in foci (protein pools) that co-localize with the Evf2 RNA cloud. Co-localization of
the Evf2-RNP with the Dlx5/6 UCE and Evf2-regulated genes, reveals important
mechanistic information about Evf2-dependent gene regulation. Intriguingly, Evf2 is not
always retained at its site of expression (the Dlx5/6 UCE) but can interact with Evf2-
regulated targets independent of the UCE, suggesting that interactions of Evf2 with the
chromosome are dynamic.
 Visualization of the Evf2-RNP at the single-cell level provides essential information
for understanding Evf2 regulatory mechanisms at spatiotemporal resolution. However,
imaging experiments utilizing fixed cells fail to reveal important information about dynamic
processes. In this proposal, we will utilize recent advances in live-cell imaging of RNAs
for the visualization of Evf2 in living cells. We will generate mouse models for the study
of Evf2 cloud formation, Evf2 cloud dynamics, and Evf2-RNP formation. This study will
answer critical questions for the advancement of knowledge about mechanisms of
transcriptional regulation by lncRNAs.

## Key facts

- **NIH application ID:** 10195705
- **Project number:** 1R03MH126145-01
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Jhumku Dutt Kohtz
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $79,917
- **Award type:** 1
- **Project period:** 2021-08-16 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10195705

## Citation

> US National Institutes of Health, RePORTER application 10195705, Mouse models for live-cell imaging of endogenous Evf2 lncRNA (1R03MH126145-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10195705. Licensed CC0.

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