Abstract
The Drosophila brain contains tens of thousands of distinct cell types. Thousands of different transgenic lines reproducibly target specific neuron subsets, yet most still express in several cell types. Furthermore, most lines were developed without a priori knowledge of where the transgenes would be expressed. To aid in the development of cell type–specific tools for neuronal identification and manipulation, we developed an iterative assay for transposase-accessible chromatin (ATAC) approach. Open chromatin regions (OCRs) enriched in neurons, compared to whole bodies, drove transgene expression preferentially in subsets of neurons. A second round of ATAC-seq from these specific neuron subsets revealed additional enriched OCR2s that further restricted transgene expression within the chosen neuron subset. This approach allows for continued refinement of transgene expression, and we used it to identify neurons relevant for sleep behavior. Furthermore, this approach is widely applicable to other cell types and to other organisms.
| Original language | English (US) |
|---|---|
| Article number | eadi4393 |
| Journal | Science Advances |
| Volume | 10 |
| Issue number | 13 |
| DOIs | |
| State | Published - Mar 2024 |
Funding
Acknowledgments: We thank members of the A.R. and A.R.R. laboratories for discussion. We acknowledge the cell imaging core at the University of Utah for use of equipment (leica SP8 White light laser confocal microscope). Stocks obtained from the Bloomington Drosophila Stock center (nih P40Od018537) were used in this study. this work was supported by the University of Utah Flow cytometry Facility, the University of Utah Genomics core Facility, the high throughput Sequencing core at the huntsman cancer institute, and the national cancer institute through award number 5P30cA042014. Funding: this work was supported by the national institute on drug Abuse Grant R21dA049635 (to A.R.); national institute on Alcohol Abuse and Alcoholism Grants K01AA029200 (to c.B.M.), R01AA026818 (to A.R.), R01AA019526 (to A.R.), and R01AA030881 (to A.R.); and national institute of diabetes and digestive and Kidney disease Grant R01dK110358 (to A.R.R.). Author contributions: conceptualization: c.B.M. and A.R. Formal analysis: c.B.M., i.t., A.B.M., and A.R. investigation: c.B.M., i.t., and M.A.P. Resources: M.A.P. data curation: c.B.M. and A.B.M. Writing\u2014original draft: c.B.M. and i.t. Writing\u2014review and editing: c.B.M., i.t., M.A.P., A.R.R., and A.R. visualization: c.B.M. Supervision: A.R. Funding acquisition: c.B.M., A.R.R., and A.R. Competing interests: the authors declare that they have no competing interests. Data and materials availability: All sequencing data are deposited in the Gene expression Omnibus database (accession number: GSe226514). All generated plasmids and fly lines are available upon request. All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials.
ASJC Scopus subject areas
- General