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| Status |
Public on Jun 29, 2020 |
| Title |
Reciprocal gene editing defines targetable mutant H3.3 oncohistone effectors in pediatric glioma |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
High-grade pediatric gliomas often contain histone H3.3 mutations, but open questions remain about oncogenic mechanisms. To address this gap, we performed ‘reciprocal gene editing’ using CRISPR-Cas9 to introduce H3.3 mutations (K27M, G34R) into H3.3-wildtype brain and glioma cells, while in parallel reverting pre-existing K27M mutations in glioma cells back to wildtype. Analyses of our reciprocally-edited cells indicate that H3.3 mutation leads to specific transcriptomic and epigenetic events, and associated cell biological changes including in xenograft assays. We used these data and the reciprocally-edited cells to screen selected drugs and identify specific putative treatments that are mutant H3.3-specific. Overall, reciprocal gene editing provides new insights into mutant H3.3 oncogenic mechanisms and more broadly may prove useful for studying other cancer-associated mutations.
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| Overall design |
We used CRISPR-Cas9 gene editing of H3F3A to precisely revert preexisting K27M mutations to WT in patient-derived glioma cells to identify consistently functionally important effectors. Two replicates of H3K27me3 and two replicates of H3.3 ChIP-Seq were performed in each of the following cell lines: Line XIII, Line XIII WT, Line XVII, Line XVII WT.
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| Contributor(s) |
Klein R, Knoepfler P |
| Citation(s) |
32647372 |
| Submission date |
Apr 12, 2019 |
| Last update date |
Jul 20, 2020 |
| Contact name |
Paul Knoepfler |
| E-mail(s) |
knoepfler@ucdavis.edu
|
| Organization name |
University of California Davis
|
| Department |
Cell Biology and Human Anatomy
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| Street address |
2425 Stockton Blvd, Room 633
|
| City |
Sacramento |
| State/province |
CA |
| ZIP/Postal code |
95817 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (20)
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| Relations |
| BioProject |
PRJNA532652 |
| SRA |
SRP192471 |
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