Peikon, I. D., Kebschull, J. M., Vagin, V. V., Ravens, D. I., Sun, Y. C., Brouzes, E., Correa, I. R., Bressan, D., Zador, A. M. (April 2017) Using high-throughput barcode sequencing to efficiently map connectomes. Nucleic Acids Res. ISSN 0305-1048
Preview |
PDF (Paper)
Zador 2017.pdf - Published Version Download (4MB) | Preview |
Abstract
The function of a neural circuit is determined by the details of its synaptic connections. At present, the only available method for determining a neural wiring diagram with single synapse precision-a 'connectome'-is based on imaging methods that are slow, labor-intensive and expensive. Here, we present SYNseq, a method for converting the connectome into a form that can exploit the speed and low cost of modern high-throughput DNA sequencing. In SYNseq, each neuron is labeled with a unique random nucleotide sequence-an RNA 'barcode'-which is targeted to the synapse using engineered proteins. Barcodes in pre- and postsynaptic neurons are then associated through protein-protein crosslinking across the synapse, extracted from the tissue, and joined into a form suitable for sequencing. Although our failure to develop an efficient barcode joining scheme precludes the widespread application of this approach, we expect that with further development SYNseq will enable tracing of complex circuits at high speed and low cost.
| Item Type: | Paper |
|---|---|
| Subjects: | organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits Investigative techniques and equipment > Whole Brain Circuit Mapping |
| CSHL Authors: | |
| Communities: | CSHL labs > Zador lab School of Biological Sciences > Publications |
| Depositing User: | Matt Covey |
| Date: | 26 April 2017 |
| Date Deposited: | 28 Apr 2017 16:30 |
| Last Modified: | 07 Jul 2021 13:53 |
| PMCID: | PMC5499584 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/34646 |
Actions (login required)
 |
Administrator's edit/view item |