| Project/Area Number |
23K14545
|
|---|
| Research Category |
Grant-in-Aid for Early-Career Scientists
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Basic Section 49070:Immunology-related
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
Chen KelvinYigene 大阪大学, 免疫学フロンティア研究センター, 特任助教(常勤) (00898851)
|
|---|
| Project Period (FY) |
2023-04-01 – 2026-03-31
|
| Project Status |
Granted (Fiscal Year 2024)
|
| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2024: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2023: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
|
|---|
| Keywords | Single cell genonmics / Immunology / Thymus development / CRISPR |
|---|
| Outline of Research at the Start |
The applicant proposes the a novel CRISPR screening platform as a powerful tool for understanding phenotypic changes of the transcriptome, epigenome and proteome that can be further extended towards other CRISPR-based applications.
|
| Outline of Annual Research Achievements |
We have now tested the related technology and novel lentiviral vectors in several in vitro contexts using primary human CD4 T cells, with good success. We achieved efficient gRNA capture and assignment in multi-modal single-cell genomic assays without any noticeable decrease in assay quality. Moreover, using these assays, we fine-mapped the underlying gene regulatory networks governing T cell activation. These assays enable scalable, multi-omic phenotypic characterization of genetic perturbations.
We have also further engineered the viral vector by incorporating virally derived sequence motifs that enhance sequence stability, reduce secondary structure, and provide additional priming sites for reverse transcription of the gRNA capture region.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The progress thus far has been proceeding as expected without too much delay.
|
| Strategy for Future Research Activity |
Now that we have a working model in vitro that has benchmarked well against the current golden standards, we are now planning on applying these technologies into in vivo settings. We will look to dissect trans-factor-mediated gene regulatory networks in the context of T cell differentiation through use of CRISPR perturbation screenings and single-cell genomics.
|
