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| Status |
Public on Aug 13, 2020 |
| Title |
Paired Imaging and Sequencing of Protein-DNA Interactions in Single Human Cells Using microDamID |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
Expression profiling by high throughput sequencing
|
| Summary |
Genome regulation depends on carefully programmed protein-DNA interactions that maintain or alter gene expression states, often by influencing chromatin organization. Most studies of these interactions to date have relied on bulk methods, which in many systems cannot capture the dynamic single-cell nature of these interactions as they modulate cell states. One method allowing for sensitive single-cell mapping of protein-DNA interactions is DNA adenine methyltransferase identification (DamID), which records a protein’s DNA-binding history by methylating adenine bases in its vicinity, then selectively amplifies and sequences these methylated regions. These interaction sites can also be visualized using fluorescent proteins that bind to methyladenines. Here we combine these imaging and sequencing technologies in an integrated microfluidic platform (µDamID) that enables single-cell isolation, imaging, and sorting, followed by DamID. We apply this system to generate paired single-cell imaging and sequencing data from a human cell line, in which we map and validate interactions between DNA and nuclear lamina proteins, providing a measure of 3D chromatin organization and broad gene regulation patterns. µDamID provides the unique ability to compare paired imaging and sequencing data for each cell and between cells, enabling the joint analysis of the nuclear localization, sequence identity, and variability of protein-DNA interactions.
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| |
|
| Overall design |
Single-cell DamID to map lamina associated domains in HEK293T cells imaged on a microfluidic device.
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| Web link |
https://0-doi-org.brum.beds.ac.uk/10.1016/j.cels.2020.08.015
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| |
|
| Contributor(s) |
Altemose N, Maslan A, Streets A |
| Citation(s) |
33099405 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R35 GM124916 |
OPTICAL, FLUIDIC, AND MOLECULAR TECHNOLOGIES FOR SINGLE-CELL OMICS |
UNIVERSITY OF CALIFORNIA BERKELEY |
Aaron Streets |
|
| Submission date |
Aug 12, 2020 |
| Last update date |
Nov 12, 2020 |
| Contact name |
Nicolas Altemose |
| Organization name |
University of California, Berkeley
|
| Department |
Bioengineering
|
| Lab |
Aaron Streets
|
| Street address |
327 Stanley Hall
|
| City |
Berkeley |
| State/province |
CA |
| ZIP/Postal code |
94720 |
| Country |
USA |
| |
|
| Platforms (3) |
| GPL21697 |
NextSeq 550 (Homo sapiens) |
| GPL22790 |
Illumina MiniSeq (Homo sapiens) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
|
| Samples (74)
|
|
| Relations |
| BioProject |
PRJNA656844 |
| SRA |
SRP277257 |
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