A scalable platform for the development of cell-type-specific viral drivers
- Harvard Medical School, United States
Abstract
Enhancers are the primary DNA regulatory elements that confer cell type specificity of gene expression. Recent studies characterizing individual enhancers have revealed their potential to direct heterologous gene expression in a highly cell-type-specific manner. However, it has not yet been possible to systematically identify and test the function of enhancers for each of the many cell types in an organism. We have developed PESCA, a scalable and generalizable method that leverages ATAC- and single-cell RNA-sequencing protocols, to characterize cell-type-specific enhancers that should enable genetic access and perturbation of gene function across mammalian cell types. Focusing on the highly heterogeneous mammalian cerebral cortex, we apply PESCA to find enhancers and generate viral reagents capable of accessing and manipulating a subset of somatostatin-expressing cortical interneurons with high specificity. This study demonstrates the utility of this platform for developing new cell-type-specific viral reagents, with significant implications for both basic and translational research.
Data availability
Sequencing data had been deposited in GEO under accession code GSE136802
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A scalable platform for the development of cell-type-specific viral driversNCBI Gene Expression Omnibus, GSE136802.
Article and author information
Author details
Michael E Greenberg
Funding
National Institutes of Health (RF1MH11408101)
- Michael E Greenberg
National Institutes of Health (5T32AG000222-23)
- Sinisa Hrvatin
National Institutes of Health (T32GM007753)
- M Aurel Nagy
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Anne E West, Duke University School of Medicine, United States
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#IS00000074-3) of Harvard Medical School. All surgery was performed under isoflurane anesthesia, and every effort was made to minimize suffering.
Version history
- Received: April 30, 2019
- Accepted: September 16, 2019
- Accepted Manuscript published: September 23, 2019 (version 1)
- Version of Record published: October 3, 2019 (version 2)
Copyright
© 2019, Hrvatin et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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A scalable platform for the development of cell-type-specific viral drivers
eLife 8:e48089.
https://doi.org/10.7554/eLife.48089
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