 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Jun 30, 2023 |
| Title |
NanoDam Profiling of EGL-43 in the Anchor Cell |
| Organism |
Caenorhabditis elegans |
| Experiment type |
Other
|
| Summary |
Depending on the cellular context, TF expression can vary dramatically both spatially and temporally. These differences in expression patterns can result in tissue-specific differences in TF binding to downstream targets. To identify targets on a tissue-specific basis, Targeted DamID (TaDa) has been recently introduced to generate TF binding profiles in various models including C. elegans. However, TaDa suffers from portability such that a new promoter-TF fusion transgene must be constructed for every new experimental condition of interest. Here, we adapt NanoDam for usage in C. elegans, which relies on the use of endogenous TF-GFP knock-ins, a plethora of which have already been generated by the community. We report that NanoDam single copy transgenes consisting of lowly expressed, tissue-specific GFP nanobody-Dam fusions, when combined with endogenous GFP-tagged alleles of TFs, results in robust, tissue-specific profiling. Using an endogenous GFP-tagged allele of EGL-43/EVI1, we performed NanoDam profiling of two disparate tissue types, the anchor cell (AC) and dopaminergic neurons, and identify targets unique to each and shared by both cell types. We also identify two GATA TFs, ELT-6 and EGL-18, as novel regulators of AC invasion. Taken together, we demonstrate that NanoDam is capable of profiling endogenous GFP-tagged TFs to identify novel downstream targets in specific cell types of C. elegans.
|
| |
|
| Overall design |
A single copy insertion of an Anchor Cell promoter driving expression of a GFP nanobody fused to Dam methylase was crossed together with animals carrying endogenous EGL-43::GFP. Dam methylase permanently methylated DNA in close proximity to regions of TF binding. DNA from L3 worms was extracted, digested with DpnI, amplified, and sequenced using the MinION Nanopore platform
|
| |
|
| Contributor(s) |
Yee C, Shen K |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
May 19, 2023 |
| Last update date |
Jun 30, 2023 |
| Contact name |
Callista Yee |
| E-mail(s) |
calyee@stanford.edu
|
| Organization name |
Stanford University/HHMI
|
| Street address |
327 Campus Drive, Shen lab, Bass Biology 329
|
| City |
Stanford |
| State/province |
California |
| ZIP/Postal code |
94305 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL26522 |
MinION (Caenorhabditis elegans) |
|
| Samples (6)
|
| GSM7397962 |
L3 animals with lin-29_NanoDam control, replicate 1 |
| GSM7397963 |
L3 animals with lin-29_NanoDam control, replicate 2 |
| GSM7397964 |
L3 animals with lin-29_NanoDam control, replicate 3 |
| GSM7397965 |
L3 animals with lin-29 NanoDam and EGL-43::GFP, replicate 1 |
| GSM7397966 |
L3 animals with lin-29 NanoDam and EGL-43::GFP, replicate 2 |
| GSM7397967 |
L3 animals with lin-29 NanoDam and EGL-43::GFP, replicate 3 |
|
| Relations |
| BioProject |
PRJNA974461 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc-FDR0.01.peaks.rep1.gff.gz |
16.9 Kb |
(ftp)(http) |
GFF |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc-FDR0.01.peaks.rep2.gff.gz |
16.0 Kb |
(ftp)(http) |
GFF |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc-FDR0.01.peaks.rep3.gff.gz |
17.1 Kb |
(ftp)(http) |
GFF |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc.rep1.bedgraph.gz |
3.4 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc.rep2.bedgraph.gz |
3.1 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE232946_lin-29_NanoDam_egl-43-vs-Dam.gatc.rep3.bedgraph.gz |
3.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE232946_lin-29_NanoDam_egl-43_combined_1500.bed.gz |
37.9 Kb |
(ftp)(http) |
BED |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
|
|
|
|
 |
Less...
More...