# Regulation and Function of Alternative mRNA Isoform Expression in Mammals

> **NIH NIH R01** · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · 2024 · $528,210

## Abstract

Summary
Most human genes contain introns, and presence of introns often increases the expression of the host gene, a
phenomenon known as intron-mediated enhancement (IME). IME has been observed in diverse genes in
animals, plants and fungi and often varies in magnitude across introns. However, little is known about how
introns impact expression or what intron features modulate IME activity. Recently, we have described a novel
phenomenon that we call exon-mediated activation of transcription starts (EMATS), in which the splicing of
internal exons impacts the spectrum of promoters used and expression level of the gene. EMATS acts at a
distance of up to a few kb, can alter gene expression by at least severalfold, and appears more active at
certain promoters – especially intrinsically weak promoters. The detailed sequence requirements and mode of
action of EMATS are not yet known. This proposal is seeks to understand the rules that govern IME and
EMATS, to improve the prediction of gene expression and to enable methods to modulate gene expression by
altering splicing. It is organized around the following aims.
SA1. Determine the sequence dependence of intron-mediated enhancement.
SA2. Explore the scope and rules for EMATS regulation.
In Aim 1, we will generate a library of many thousands of distinct random sequences inserted into an intron in a
dual fluorescent reporter system that is chromosomally integrated into human cells. This design will enable
high-throughput measurement of the effects of each intron on nascent RNA, mature RNA and protein levels,
and these data will be used to identify motifs that enhance or silence expression in a splicing-dependent
manner from an intronic location. In aim 2, we will systematically derive and test rules for how EMATS
regulation depends on the location and sequence of the internal exon and on properties of the involved
promoter. Finally, we will use the information learned about IME and EMATS to improve predictions of gene
expression from primary sequence. Together, the research described in these aims will establish rules
governing how splicing impacts gene expression in mammalian genomes. Identification of motifs that function
as splicing-dependent activators or silencers of expression can be used to improve prediction of expression
from genome sequence and may enable detection of intronic variants that alter expression. Understanding
how splicing impacts expression may also enable new approaches for gene expression modulation.

## Key facts

- **NIH application ID:** 10761767
- **Project number:** 5R01HG002439-20
- **Recipient organization:** MASSACHUSETTS INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** CHRISTOPHER B BURGE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $528,210
- **Award type:** 5
- **Project period:** 2021-01-06 → 2025-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10761767

## Citation

> US National Institutes of Health, RePORTER application 10761767, Regulation and Function of Alternative mRNA Isoform Expression in Mammals (5R01HG002439-20). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10761767. Licensed CC0.

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