# Generation of viral vectors that use alternative splicing to drive cell type-specific gene expression in the nervous system

> **NIH NIH RF1** · JOHNS HOPKINS UNIVERSITY · 2020 · $1,534,482

## Abstract

Project Summary
New tools are urgently needed to selectively target constructs that monitor and manipulate the activity of
individual cell types without having to rely on genetic manipulation. This proposal aims to develop viral tools
that use cell type-specific alternative splicing events to drive cell type-specific gene expression in the nervous
system independent of genetic manipulation, an approach we term splicing-linked expression design (SLED).
SLED-based vectors use evolutionarily conserved, highly cell type-specific exons, identified using the ASCOT
database developed by our group, to drive expression of reporter and effector constructs. We have
demonstrated feasibility of this approach using constructs that selectively target retinal photoreceptors, muscle
cells, and cortical neurons. We propose to extend this by combining cell-specific alternative exon/intron
sequences with appropriate promoter sequences to generate a toolbox of AAV and lentiviral SLED vectors that
selectively target multiple cell types of interest to the neuroscience research community. We will first generate
SLED vectors that target primary sensory and motor neurons, as well as multiple subtypes of cortical neurons
and glia, and validate the specificity of these reagents in mice. We will next test the cell specificity of SLED
reagents that are validated in mice in rats, ferrets, as well as human cortical organoids and rat-human
chimeras. Finally, highly specific SLED fluorescent reporter constructs will be converted to drive expression of
calcium indicators, as well as optogenetic and chemogenetic constructs. We anticipate that SLED-based
reagents will allow highly cell type-specific expression of a broad range of molecular tools useful for analysis of
neural circuitry in multiple mammalian species.

## Key facts

- **NIH application ID:** 10012468
- **Project number:** 1RF1MH123237-01
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Seth Blackshaw
- **Activity code:** RF1 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $1,534,482
- **Award type:** 1
- **Project period:** 2020-08-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10012468

## Citation

> US National Institutes of Health, RePORTER application 10012468, Generation of viral vectors that use alternative splicing to drive cell type-specific gene expression in the nervous system (1RF1MH123237-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10012468. Licensed CC0.

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