Experiment Overview
| Repository ID: | FR-FCM-Z2LQ | Experiment name: | Engineering monocyte/macrophage specific glucocerebrosidase expression in human hematopoietic stem cells using genome editing | MIFlowCyt score: | 60.22% |
| Primary researcher: | Samantha Scharenberg | PI/manager: | Samantha Scharenberg | Uploaded by: | Samantha Scharenberg |
| Experiment dates: | 2016-06-20 - 2020-04-15 | Dataset uploaded: | May 2020 | Last updated: | May 2020 |
| Keywords: | [CRISPR] [HSC] [Gaucher] [Gene-editing] | Manuscripts: | |||
| Organizations: |
Stanford University, Baxter Laboratory for Stem Cell Biology, Stanford, CA (United States)
Stanford University - Gomez-Ospina Lab, Pediatrics, Palo Alto, (CA) |
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| Purpose: | Determine the feasibility of using CRISPR/Cas9 and AAV to genetically engineer human hematopoietic stem/progenitor cells (HSPCs) to express glucocerebrosidase under a macrophage-specific promoter as a curative therapy for Gaucher Disease. | ||||
| Conclusion: | CRISPR/Cas9 and AAV achieve efficient targeted integration of human glucocerebrosdiase-expressing cassettes into human HSPCs. Targeted HSPCs exhibit mcarophage-specific cassette expression and are capable of long-term multi-lineage engraftment (including tissue-resident macrophages) in the NSG murine model . | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Negative controls (such as mock-treated cells) were used to draw marker-positive gates; cells treated with AAV vector alone (-RNP) were used to determine background episomal vector expression; compensation controls and FMOS were used where appropriate | ||||
Experiment variables
| Timepoints | |
|---|---|
| · Day 0 | Figure 1d_D2_CD68S AAV+RNP.fcs · Figure 1d_D2_Mock.fcs · Figure 1d_D2_SFFV AAV only.fcs · Figure 1d_D2_SFFV AAV+RNP.fcs · Figure 1d_D3_CD68S AAV only.fcs · Figure 1d_D3_CD68S AAV only.fcs-(1) · Figure 1d_D3_CD68S AAV+RNP.fcs · Figure 1d_D3_Mock.fcs · Figure 1d_D3_SFFV AAV only.fcs · Figure 1d_D3_SFFV AAV+RNP.fcs |
| · Day 4 | Figure 3 bc_D1_D4_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D1_D4_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D1_D4_LTC_Mock.fcs · Figure 3 bc_D1_D4_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D1_D4_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D1_D4_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D1_D4_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D1_D4_Mphage_Mock.fcs · Figure 3 bc_D1_D4_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D1_D4_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D2_D4_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D2_D4_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D2_D4_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D2_D4_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D2_D4_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D2_D4_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D2_D4_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D2_D4_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D3_D4_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D3_D4_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D3_D4_LTC_Mock.fcs · Figure 3 bc_D3_D4_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D3_D4_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D3_D4_Mphage_CD68S-Citrine+fcs · Figure 3 bc_D3_D4_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D3_D4_Mphage_Mock.fcs · Figure 3 bc_D3_D4_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D3_D4_Mphage_SFFV-Citrine-.fcs |
| · Day 8 | Figure 3 bc_D1_D8_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D1_D8_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D1_D8_LTC_Mock.fcs · Figure 3 bc_D1_D8_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D1_D8_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D1_D8_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D1_D8_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D1_D8_Mphage_Mock .fcs · Figure 3 bc_D1_D8_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D1_D8_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D2_D8_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D2_D8_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D2_D8_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D2_D8_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D2_D8_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D2_D8_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D2_D8_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D2_D8_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D3_D8_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D3_D8_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D3_D8_LTC_Mock.fcs · Figure 3 bc_D3_D8_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D3_D8_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D3_D8_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D3_D8_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D3_D8_Mphage_Mock.fcs · Figure 3 bc_D3_D8_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D3_D8_Mphage_SFFV-Citrine-.fcs |
| · Day 12 | Figure 3 bc_D1_D12_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D1_D12_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D1_D12_LTC_Mock.fcs · Figure 3 bc_D1_D12_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D1_D12_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D1_D12_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D1_D12_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D1_D12_Mphage_Mock.fcs · Figure 3 bc_D1_D12_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D1_D12_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D2_D12_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D2_D12_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D2_D12_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D2_D12_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D2_D12_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D2_D12_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D2_D12_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D2_D12_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D3_D12_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D3_D12_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D3_D12_LTC_Mock.fcs · Figure 3 bc_D3_D12_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D3_D12_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D3_D12_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D3_D12_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D3_D12_Mphage_Mock.fcs · Figure 3 bc_D3_D12_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D3_D12_Mphage_SFFV-Citrine-.fcs |
| · Day 16 | Figure 3 bc_D1_D16_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D1_D16_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D1_D16_LTC_Mock.fcs · Figure 3 bc_D1_D16_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D1_D16_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D1_D16_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D1_D16_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D1_D16_Mphage_Mock.fcs · Figure 3 bc_D1_D16_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D1_D16_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D2_D16_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D2_D16_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D2_D16_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D2_D16_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D2_D16_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D2_D16_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D2_D16_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D2_D16_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D3_D16_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D3_D16_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D3_D16_LTC_Mock.fcs · Figure 3 bc_D3_D16_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D3_D16_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D3_D16_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D3_D16_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D3_D16_Mphage_Mock.fcs · Figure 3 bc_D3_D16_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D3_D16_Mphage_SFFV-Citrine-.fcs |
| · Day 20 | Figure 3 bc_D1_D20_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D1_D20_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D1_D20_LTC_Mock.fcs · Figure 3 bc_D1_D20_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D1_D20_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D1_D20_Mphage Adherent_CD68S-Citrine+.fcs · Figure 3 bc_D1_D20_Mphage Adherent_CD68S-Citrine-.fcs · Figure 3 bc_D1_D20_Mphage Adherent_SFFV-Citrine+.fcs · Figure 3 bc_D1_D20_Mphage Adherent_SFFV-Citrine-.fcs · Figure 3 bc_D1_D20_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D1_D20_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D1_D20_Mphage_Mock.fcs · Figure 3 bc_D1_D20_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D1_D20_Mphage_SFFV-Citrine-.fcs · Figure 3 bc_D2_D20_LTC_CD68S-Citrine+.fcs · Figure 3 bc_D2_D20_LTC_CD68S-Citrine-.fcs · Figure 3 bc_D2_D20_LTC_SFFV-Citrine+.fcs · Figure 3 bc_D2_D20_LTC_SFFV-Citrine-.fcs · Figure 3 bc_D2_D20_Mphage Adherent_CD68S-Citrine+.fcs · Figure 3 bc_D2_D20_Mphage Adherent_CD68S-Citrine-.fcs · Figure 3 bc_D2_D20_Mphage Adherent_SFFV-Citrine+.fcs · Figure 3 bc_D2_D20_Mphage Adherent_SFFV-Citrine-.fcs · Figure 3 bc_D2_D20_Mphage_CD68S-Citrine+.fcs · Figure 3 bc_D2_D20_Mphage_CD68S-Citrine-.fcs · Figure 3 bc_D2_D20_Mphage_SFFV-Citrine+.fcs · Figure 3 bc_D2_D20_Mphage_SFFV-Citrine-.fcs |
