Coupling single cell RNA sequencing with crispr perturbation and using single cell RNA sequencing to profile musculoskeletal injuries and disease
Abstract/Contents
- Abstract
- Single cell RNA sequencing (scRNAseq) has changed the way we understand cellular heterogeneity and overall tissue behavior. Using scRNAseq, we addressed three questions here, one technical, two biological. For the first question, we wanted to assess if Clustered regularly interspaced short palindromic repeats (CRISPR) perturbation information can be coupled with transcriptomic information in the drop-seq setting through direct capture of the small guide RNA (sgRNA). While capturing sgRNAs on drop-seq beads was achieved, we found that our downstream signal was scrambled. The first biological question we addressed was looking at an Hsl knockout mouse with faster fracture repair. Initial experiments suggested osteoimmunological causes behind the faster repair. Using scRNAseq, we found that pre-osteoclasts were the most perturbed cell type suggesting they may play a role in the improved fracture repair. Furthermore, pre-osteoclasts were some of the few cells in our dataset that expressed Pparg for which Hsl makes ligands. The second biological question we addressed was looking at tendon progenitor cells (TPCs) derived from either tendinopathic or normal patellar tendon. In our mechanical stress experimental system we found substantial differences between normal and tendinopathy TPCs. This dissertation illustrates the potential of scRNAseq applications to address a number of important biological questions while still have room to be further expanded.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Still, Christopher Darryl |
|---|---|
| Degree supervisor | Qi, Lei, (Professor of Bioengineering) |
| Thesis advisor | Qi, Lei, (Professor of Bioengineering) |
| Thesis advisor | Alizadeh, Ash |
| Thesis advisor | Oro, Anthony, 1958- |
| Thesis advisor | Wu, Joy |
| Degree committee member | Alizadeh, Ash |
| Degree committee member | Oro, Anthony, 1958- |
| Degree committee member | Wu, Joy |
| Associated with | Stanford University, Department of Stem Cell Biology and Regenerative Medicine |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Christopher Darryl II. |
|---|---|
| Note | Submitted to the Department of Stem Cell Biology and Regenerative Medicine. |
| Thesis | Thesis Ph.D. Stanford University 2020. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Christopher Darryl Still
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
Also listed in
Loading usage metrics...